Compositions and methods for maintaining or enhancing homeostasis or function of female lower reproductive tract

ABSTRACT

The present disclosure provides compositions and methods for making and using topical compositions comprised of an isotonic, biome-friendly solution containing xylose and/or a  Salvia  extract. The compositions are useful for, for example, maintaining or enhancing female lower reproductive tract (LRT) homeostasis and physiological function. In particular, topical compositions of this disclosure can be formulated for use as a freshening, menopausal, fertility, and/or perineal composition.

BACKGROUND

Mounting evidence indicates that cells lining the lower femalereproductive tract (LRT), including that of the vulva, vagina andexternal cervix, together with the resident commensal, vaginotropicmicrobiome (collectively, the “vaginal ecosystem”), form a dynamic organthat not only protects against infectious disease but also maintainstissue health and facilitates reproduction. The vaginal ecosystem ismade up of a variety of factors that together act to support homeostasisof this ecosystem, including immune active cells, a protective mucuscoating, a variety of antimicrobial peptides, and symbiotic bacteria(e.g., Lactobacilli).

To maintain homeostasis, the vaginal ecosystem must adapt to changesthat occur during a woman's monthly hormone cycle, as well as adjust tomore profound changes that happen at different times in the arc of awoman's life, such as puberty, reproductive years, and menopause. Forexample, during various stages of a woman's reproductive cycle, thevaginal ecosystem is continually providing a protective barrier todisease while at the same time lubricating the vagina to accommodate forsexual intercourse, or facilitating the passage of blood out of theuterus at menses, or inhibiting/optimizing sperm transport through thevagina and cervix, or undergoing chemical changes to promote labor andbirth of a baby.

Throughout a woman's life, the LRT and vaginal ecosystem continuallyexperience various disturbances (acute and chronic) caused by humanbehavior (e.g., sexual intercourse), but also by the use ofcontraception and vaginal products (Hickey et al., Transl. Res. 160:267,2012; Ma et al., Annu. Rev. Microbiol. 66:371, 2012). If disturbance ofthe vaginal ecosystem becomes chronic, then discomfort, and evendisease, can ensue. Any vaginal product that alters the mucosalenvironment and impairs the epithelial barrier may increase the severityof symptoms or risk of disease (Fichorova et al., Toxicol. Appl.Pharmacol. 285:198, 2015; Ma et al., 2012).

The consequence of products for women's health that ignores the fullbiology of the LRT and the vaginal ecosystem has been costly at both theepidemiologic and personal level. Vaginal products developed to preventSexually Transmitted Diseases (STDs) were actually found to cause anincrease in STD rates when tested in clinical trials (Gali et al.,Antimicrob. Agents Chemother. 54:5105, 2010; Dezzutti et al., PLoS One7:e48328, 2012). Cell toxicity of various ingredients (some are referredto as “inactive carriers”) in vaginally applied products, for both thehealthy vaginotropic bacteria and the vaginal mucosal epithelial cells,is pervasive and is evident at concentrations below those found incommon vaginal products.

Safer LRT product compositions to aid in women's health are needed, suchas products designed to preserve and support the vaginal ecosystemhomeostasis and healthy function thereof. Presently disclosedembodiments address this need and provide other related advantages.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a bar graph depicting the effect on sperm when contacted withan isotonic, neutral pH (pH 7) solution with 10% Salvia hispanicaextract (in an oil format) as compared to canola oil and almond oil. ASperm Toxicity Index of <0.75 is considered cytotoxic to the sperm.

FIG. 2 is a bar graph depicting mucosal irritation potential as measuredusing slug mucosal irritation index for Fertility Gel 1a and FresheningGel 2a, isotonic solutions containing xylose, as compared with FertilityGel 1b (also referred to as Fertility Gel Formula AB) and Freshening Gel2b (also referred to as Freshening Gel Formula AB), in which Salviasclarea extract has been added.

FIG. 3 is a bar graph depicting the mucosal irritation potential asmeasured by mucus production using the slug mucosal irritation index forFreshening Gel 2b, a pH balancing and freshening gel, as compared withMONISTAT™ and REPHRESH™.

FIG. 4 is a bar graph depicting the effect of Freshening Gel 2b on spermmotility, as compared with MONISTAT™ and REPHRESH™.

FIG. 5 is a bar graph depicting the mucosal irritation potential asmeasured using slug mucosal irritation index for Fertility Gel 1b, aformulation for enhanced sperm function during ovulatory stage of thefemale menstrual cycle as compared to a paraben-containing commercialfertility gel and KY Jelly®.

FIG. 6 is a bar graph depicting percent sperm motility of semen samplesincubated for 30 minutes with 10% solution of Fertility Gel 1c ascompared to a control.

FIG. 7 is a bar graph depicting total motile sperm concentration ofsemen samples incubated for 30 minutes with 10% solution of FertilityGel 1c as compared to a control.

FIG. 8 is a bar graph depicting % DNA fragmentation of sperm chromatinfrom semen samples incubated with Fertility Gel 1b as measured by thesperm chromatin structure assay (SCSA).

FIGS. 9A and 9B: FIG. 9A depicts a 10 cm visual analog scale (not drawnto scale) for evaluating slipperiness of Fertility Gel 1c and aparaben-containing commercial fertility gel. FIG. 9B is a bar graphdepicting the degree of slipperiness Fertility Gel 1c and aparaben-containing commercial fertility gel as measured by the distance(cm) of the participants marking on the visual analog scale.

DETAILED DESCRIPTION

The present disclosure provides compositions and methods for maintainingor enhancing homeostasis, function or both of the female lowerreproductive tract (LRT). In particular, the instant disclosure relatesto unique isotonic, biome-friendly formulations comprising xylose and aSalvia extract at various pH levels to complement unique hormone cycleand life-stage specific physiology and function of the female LRT. Suchisotonic, biome-friendly formulations of xylose and Salvia are usefulfor prophylactically or therapeutically promoting and maintaining anatural balance in the female LRT environment, including the vaginalecosystem. More specifically, isotonic, biome-friendly formulations ofthis disclosure can be used for treating or preventing infection,preventing conception, enhancing fertility, maintaining or enhancingvaginal ecosystem homeostasis, increasing hydration of the vaginalmucosa, or the like.

Prior to setting forth this disclosure in more detail, it may be helpfulto an understanding thereof to provide definitions of certain terms tobe used herein. Additional definitions are set forth throughout thisdisclosure.

In the present description, any concentration range, percentage range,ratio range, or integer range is to be understood to include the valueof any integer within the recited range and, when appropriate, fractionsthereof (such as one tenth and one hundredth of an integer), unlessotherwise indicated. Also, any number range recited herein relating toany physical feature, such as polymer subunits, size or thickness, areto be understood to include any integer within the recited range, unlessotherwise indicated. Any concentration ranges recited herein are to beunderstood to include concentrations of any integer within the range andfractions thereof, such as one tenth, one hundredth, and one thousandthof an integer, unless otherwise indicated. As used herein, the term“about” means±20% of the indicated range, value, or structure, unlessotherwise indicated. The term “consisting essentially of” limits thescope of a claim to the specified materials or steps, or to those thatdo not materially affect the basic and novel characteristics of theclaimed invention. It should be understood that the terms “a” and “an”as used herein refer to “one or more” of the enumerated components. Theuse of the alternative (e.g., “or”) should be understood to mean eitherone, both, or any combination thereof of the alternatives. As usedherein, the terms “include,” “have” and “comprise” are usedsynonymously, which terms and variants thereof are intended to beconstrued as non-limiting.

A “pharmaceutical composition” or “vaginal composition” refers to aformulation of this disclosure and a medium generally accepted in theart for the delivery of the biologically effective compound(s) to femalesubjects, e.g., humans. For example, a pharmaceutical or vaginalcomposition of the present disclosure may be formulated or used as astand-alone composition, or as a component in a prescription drug, anover-the-counter (OTC) medicine, a botanical drug, an herbal medicine, amedical device, a cosmetic, a homeopathic agent, or any other form ofhealth care product, optionally reviewed and approved by a governmentagency. A medium generally accepted in the art includes allpharmaceutically or vaginally acceptable carriers, diluents orexcipients therefor.

As used herein, “enriched for” refers to a plant extract or otherpreparation having at least a two-fold up to about a 1000-fold increaseof one or more active compounds as compared to the amount of one or moreactive compounds found in the weight of the plant material or othersource before extraction or other preparation. In certain embodiments,the weight of the plant material or other source before extraction orother preparation may be dry weight, wet weight, or a combinationthereof.

As used herein, “purified” refers to a plant extract or otherpreparation that has at least one contaminant removed therefrom or theremoval of a desired component from the milieu. In certain embodiments,the plant extract is purified using a method selected from distillation,recrystallization, precipitation, solvent partition, chromatographicseparation, or other suitable methods.

As used herein, “major active ingredient” or “major active component”refers to one or more active compounds found in a plant extract or otherpreparation, or enriched for in a plant extract or other preparation,which is capable of at least one biological activity. In certainembodiments, a major active ingredient of an enriched extract will bethe one or more active compounds that were enriched in that extract.Generally, one or more major active components will impart, directly orindirectly, most (i.e., greater than 50%) of one or more measurablebiological activities or effects as compared to other extractcomponents. In certain embodiments, a major active ingredient may be aminor component by weight percentage of an extract (e.g., less than 50%,25%, or 10% of the components contained in an extract) but still providemost of the desired biological activity. Any composition of thisdisclosure containing a major active ingredient may also contain minoractive ingredients that may or may not contribute to the pharmaceuticalor vaginal activity of the enriched composition, but not to the level ofmajor active components, and minor active components alone may not beeffective in the absence of a major active ingredient.

“Effective amount” or “therapeutically effective amount” refers to thatamount of a composition of this disclosure which, when administered to afemale mammal, such as a human, is sufficient to effect a desiredbiological effect or treatment, including any one or more of: (1)improving or promoting perineal elasticity to lessen trauma or tearingof the perineum during birth; (2) maintaining, supporting or promotingvaginal homeostasis; (3) providing contraceptive activity that impairssperm function and penetration; (4) promoting fertility by maintainingor improving sperm survival, function, or penetration into cervicalmucus; (5) hydrating and lubricating mucosal (vaginal) surface inreproductive-aged women or post-menopausal women; (6) protecting,cleaning, balancing or restoring feminine freshness in the vaginal area;(7) toning vulvar and vaginal tissues; and (8) providing a muco-adhesivecarrier for medications for adherence to and retention by the vaginalwall. The amount of a composition of this disclosure that constitutes an“effective amount” will vary depending on the formulation, the conditionbeing treated and its severity, the manner of administration, theduration of treatment, or the age of the subject to be treated, but canbe determined routinely by one of ordinary skill in the art havingregard to his own knowledge and to this disclosure.

“Lower Reproductive Tract (LRT) compositions” refer to combinations ofingredients of this disclosure that improve, promote, increase, manage,control, maintain, optimize, modify, reduce, inhibit, or prevent aparticular condition associated with the female vaginal ecosystem. Forexample, with regard to reproductive-related activities, LRT supplementsmay be used to promote hydration, lubrication, fertility, vaginalmicrobiome balance, or the like. Exemplary LRT compositions includexylose and a Salvia extract, optionally containing one or moreadditional ingredients for use in the vaginal ecosystem, such as aviscosity-increasing agent, a surfactant, a buffering agent, anosmolality adjuster, a pH modifier, a solvent, a preservative (e.g.,paraben-free), a humectant, or any other substance useful forsupplementing, maintaining or improving LRT homeostasis.

The “vaginal microbiome” or “vaginal biome” refers to the vaginalmicrobiota, as well as the products of the vaginal microbiota and thehost LRT environment.

“Biome-friendly” when used in reference to LRT compositions means thatthe compositions of this disclosure that are contacted with the femaleLRT will exhibit minimal mucosal irritation, inflammation orimmunotoxicity, as well as provide a supportive environment for vaginalmicrobiota.

The LRT compositions of the present disclosure may be administered to amammalian female or a male subject, including humans, bovine, canine,feline, equine, porcine, ovine, avian, rodent, lagomorph, caprine,non-human primate, or other exotic species.

Female Lower Reproductive Tract (LRT)

A woman's lower reproductive tract (LRT) refers to the vulva (includingthe perineal tissues), the vagina, and the ectocervix (i.e., thatportion of the cervix visible inside the vagina covered by squamousepithelium). The vagina and ectocervix that comprise the birth canal arecovered with non-keratinized stratified squamous epithelium, includingnumerous epithelial anchored mucin producing cells. Aspects of thephysiology of this mucous membrane are unique to the LRT. The LRT isthat portion of the female reproductive anatomy that can be contactedwith lotions, creams, gels, douches, films, powders, sprays, or thelike, which are included in products marketed for female hygiene andhealth care. These products can be in the form of cosmetics, medicaldevices or drugs, which are used for personal hygiene, contraception,lubrication, hydration, childbirth, assisted reproduction, diseasetreatment or aesthetics.

The vulva (the external genital organs) includes the mons pubis, labiamajora, labia minora, Bartholin glands, and clitoris. The externalgenital organs have three primary functions: enabling sperm to enter thebody, protecting the internal genital organs from infection, andproviding sexual pleasure. The mons pubis and labia majora are linedwith skin, mucus producing glands, and sweat and sebaceous glands. Thelabia minora is lined with a moist mucosal membrane, which is kepthydrated, similar to the vagina, through mucus secreting cells.Bartholin glands also secrete mucus for lubrication during intercourse.

LRT Homeostasis and Function

Homeostasis and function of the LRT, including the surface of the vulvarskin, the extracellular milieu of the vulva and vagina, and the vaginalcanal, is affected by a variety of factors, including ecosystem pH,composition of sugar-protein complex (glycoproteins and mucins), innateand adaptive immune system factors and cells, the normal microbiome, thepresence or absence of pathogens, systemic and local hormone levels,environmental factors, and the like.

A woman's LRT and, in particular, the vaginal ecosystem, has aparticular pH and comprises a diverse population of cell types,including epithelial cells, immune cells, vaginal microbiota, and attimes, a partner's sperm cells and invading pathogens from the externalworld. One important evolutionary aspect of a woman's LRT is the need toallow sperm penetration into the higher reproductive tract forprocreation, while at the same time limiting invading pathogens,especially the highly mutually evolved sexually transmitted diseases(STDs) that can also be present with semen.

As used herein, the term “vaginal microbiota or “VMB,” also referred toas “vaginal flora,” refers to the collective microorganisms thatnormally colonize the vulva and vagina and are non-pathogenic. Ingeneral, the VMB is primarily comprised of different strains ofLactobacillus (or related acid-producing bacterial types), which producelactic acid to keep the vaginal ecosystem as a tightly controlled acidicenvironment during much of a woman's monthly cycle in reproductive agedwomen (pH of about 3.5-5.5). Exemplary VMB species include Lactobacillusacidophilus, Lactobacillus jensenii, Lactobacillus gasseri,Lactobacillus finers, Lactobacillus crispatus, Lactobacillus plantarum,Lactobacillus fermentum, Lactobacillus brevis, Lactobacillus casei,Lactobacillus delbrueckii, Lactobacillus vaginalis, Lactobacillussalivarius. Other lactic-acid producing bacteria that may be part of theVMB include species of Atopobium, Leptotrichia, Leuconostoc,Megasphaera, Pediococcus, Streptococcus and Weissella. The species foundin normal VMB can differ between ethnic groups. Lactobacilli have beenshown to inhibit in vitro growth of pathogenic microorganisms, e.g.,Bacteroides fragilis, Escherichia coli, Gardnerella vaginalis,Mobiluncus spp., Neisseria gonorrhoeae, Peptostreptococcus anaerobius,Prevotella bivia and Staphylococcus aureus. This inhibitory action isthought to be achieved mainly through the action of lactic acid (Graverand Wade, Ann. Clin. Microbiol. Antimicrob. 10:8, 2011; Matu et al.,Anaerobe 16:210, 2010; Skarin and Sylwan, APMIS 94:399, 1986; Strus etal., J. Reprod. Med. 47:41, 2002).

Furthermore, lactobacilli may help prevent pathogenic organisms fromadhering to vaginal epithelial cells (Boris and Barbes, Microb. Infect.2:543, 2000). Other inhibitory mechanisms of lactobacilli includeproduction of hydrogen peroxide and bacteriocins (Martin and Suarez,Appl. Environ. Microbiol. 76:400, 2010; Aroutcheva et al., Am. J.Obstet. Gynecol. 185:375, 2001). While low pH is one factor in LRThomeostasis, the LRT of healthy women is not always maintained at a lowpH. The vaginal environment is episodically exposed to naturalsecretions with a neutral pH (about 7). More specifically, this exposureincludes blood flow from the uterus during menses; semen from a partnerafter sexual intercourse; and fertile cervical mucus production duringovulation. These cyclical disruptions in vaginal pH are transient andrapidly accommodated with a return to an acidic pH level in women withhealthy LRT function. Before puberty, during pregnancy and aftermenopause, a healthy pH of a woman's LRT physiologically rests at a pHof about 4.5 to about 6.8. In short, simple maintenance of an acidic pHdoes not define LRT homeostasis, but rather a healthy LRT will maintainhomeostasis and function while undergoing normal variations in pHdepending on a woman's hormone cycle and stage of life, among otherfactors.

Most healthy women have a stable VMB throughout their lives, but awoman's VMB can change depending on what reproductive stage of life sheis in, e.g., pregnancy, puberty, and menopause. The vaginotropic,“healthy” VMB is in large part derived from the woman's mother duringher birth. Over 112 different strains of lactobacillus and relatedcommensal bacteria have been identified in the vagina. Disruptions inthe VMB and vaginal ecosystem can cause numerous serious diseases, suchas bacterial vaginosis, vaginitis, postpartum infections, increased STDrates (including HIV), and pre-term labor (van de Wijgert et al., PLoSOne 9:e105998, 2014). In short, a stable VMB in women contributes to themaintenance of LRT homeostasis, which has significant public healthimportance.

Another contributor to the homeostasis and function of the LRT are localproteoglycans (PG) and glycosaminoglycans (GAGs). PGs are a majorcomponent of the human extracellular matrix, the area existing betweencells and at cell surfaces in an organism. Individual functions of PGscan be attributed to the GAG molecules attached to them. GAGs are longunbranched polysaccharides containing a repeating disaccharide unit.GAGs create viscous environments wherever they are present, which isprimarily on the surface of cells, in the extracellular matrix or insecretory vesicles of some types of cells. Natural lubricating gelsubstances produced by the human body are composed of GAGs and areinvolved in a variety of physiological functions, including skinhydration, joint function, sexual function, and reproduction. The typeand quantity of GAGs and PGs in female reproductive tissues changesbased on hormonal fluctuations and stage of life.

Assembly of functional GAGs is initiated in animal cells by transferringa xylose residue to specific serine residues in a PG core protein(Lugemwa et al., J. Biol. Chem. 271:19159, 1996). Additional sugar unitsattach to the protein PG core through the xylose sugar residue to form achain, which eventually folds into a unique biologically activesubstance. Interestingly, different combinations of single sugars and/orGAG fragments in solution cannot mimic the biological impact of thesesame GAG combinations when attached to a xylose molecule on a PGbackbone to form a large folded molecule (Tran et al., ACS Chem. Biol.8:949, 2013). Attachment to the PG core through a xylose molecule allowsthe carbohydrates in the GAG to fold into their biologically functionalshape.

Along with ensuring high viscosity of body fluids (e.g., cervical mucus,semen), GAGs provide structural integrity for passageways between cells,allowing for cell migration, such as sperm transport. The specific GAGsof physiological significance are hyaluronic acid (HA), dermatansulfate, chondroitin sulfate, heparin/heparan sulfate, and keratansulfate. GAGs are important in many aspects of reproduction.

Xylose concentration in tissues and the concentration of thexylosyltransferase (XT) enzyme, which is the first enzyme involved insugar chain attachment to the PG backbone) are highly correlated to thelevel of functional GAGs produced. The XT enzyme regulates thebiosynthesis of sugar chains attaching to the PG, and thereby thefunctionally present GAGs. The XT activity in body fluids is anindicator of the current PG biosynthesis rate. This is because XTcatalyzes the initial and rate limiting steps of attaching thecarbohydrate sugars to the protein PG backbone at the xylose residue(Muller et al., 2013, Glycoconj. J. 30:237-45). Xylose levels andxylosyltransferase activity has been shown in vivo and in vitro studiesto be regulators of GAG biosynthesis (Nadanaka & Kitagawa, Matrix Biol.35:18, 2014), the level of which is considered important for maintainingtissue homeostasis under normal and pathological conditions. Altered XTactivity in the human body also correlates with various chronic diseases(Kuhn et al., Biochem. Biophys. Res. Commun. 459:469, 2015), where mucusand mucin production and tissue health are compromised.

The biological function of PG and GAGs can also be observed in thereproductive tract where these factors preserve sperm motility and thevelocity at which the sperm swim, provide arousal fluids to facilitatesexual intercourse, as well as provide cervical mucus (CM) productionthroughout different stages of reproduction.

The contribution of PG and GAG fluctuations in the LRT homeostasis andfunction is also evident in the changes in the type of GAG seen in thecervix prior to birth, as a part of cervical “ripening” (Fischer et al.,J. Soc. Gynecol. Investig. 8:277, 2001). The molecular weight of GAGschange at specific stages of labor as compared to other times (Obara etal., Acta Obstet. Gynecol. Scand. 80:492, 2001). These changes appear torelate to epithelial barrier protection of the LRT. Specifically,females with GAG depletion in the cervix and vagina during pregnancyhave increased mucosal permeability and a “striking” increase in pretermbirths. GAGs in the vaginal canal and cervical mucus help maintain amucosal barrier to limit pathogen infiltration (Akgul et al.,Endocrinol. 153:3493, 2012; Akgul et al., J. Clin. Invest. 124:5481,2014; Ulrich et al., PLoS One 9:e104972, 2014). Factors that disruptthis highly tuned physiology of PG and GAG in the LRT will also affectLRT homeostasis, which can be harmful to the woman.

Inflammation can reduce GAG formation and chain length. When cartilagecells are challenged in the laboratory with interleukin (IL) cytokines,total xylose residues in the PGs are reduced, thus limiting theopportunity for beneficial lubricating GAGs to form (Frankenberger etal., Connect. Tissue Res. 54:123, 2013).

Iyibozkurt et al. (Reprod. Biomed Online. 19:784, 2009) teach thatchanges in GAG physiology can change cellular function. For example,vascular endothelial growth factor (VEGF) improves sperm motility, butnot sperm viability. The GAG-binding domain of VEGF has no significantsequence or structural similarity to any other known proteins and, thus,represents a novel heparin-binding domain (Fairbrother et al., Structure6:637, 1998), which could be involved in enhancing sperm function.

Mucins are another form of glycosylated protein that can impact LRTphysiology and homeostasis. Mucins are O-linked glycoproteins producedby cells that establish a physical barrier against undesirable entry offoreign materials across epithelial surfaces. They are ubiquitous inmucous secretions on cell surfaces and in body fluids (Brockhausen etal., 2009, O-GalNAc Glycans. In: Varki A, Cummings R D, Esko J D, FreezeH H, Stanley P, Bertozzi C R, Hart G W, Etzler M E, editors. Essentialsof Glycobiology. 2^(nd) ed. Cold Spring Harbor (N.Y.): Cold SpringHarbor Laboratory Press; Chapter 9) and are either secreted or membranebound (Gosalia et al., J. Biol. Chem. 288:6717, 2013).

Mucins function to hydrate and lubricate epithelial tissues(Andrianifahanana et al., Biochim. Biophys. Acta 1765:189, 2006), andthey also function in cell renewal, transport, signaling and adhesion.Mucin homeostasis is maintained by elaborate coordinated regulatorymechanisms providing a well-defined specific distribution. But, mucinhomeostasis can be disrupted by environmental or intrinsic factors.Changed gene activation can be triggered by cytokine inflammatoryagents. Resulting mis-expression can change the mucus quality beingproduced. Deregulated mucin is associated with many types of cancer andinflammatory disease. The genes that control mucin production form acluster that balances the different types of mucin products present. Incancer and during disease exposure, this mucin profile is changed.Increase in bacterial lipopolysaccharide (LPS) activates mucin geneexpression. For example, LPS at 200 ng/ml induced a 50% increase inseveral mucins and 25% decrease in others (Gosalia et al., J. Biol.Chem. 288:6717, 2013). Non-LPS mediated inflammation can also result inmucin changes. For example, inflammatory mediators in the tears of “dryeye” patients change the eye surface mucins, making them lesslubricating. (Albertsmeyer et al., 2010, Exp. Eye Res. 90:444-451). Manydiseases involve abnormal mucin secretion. Normal mucin expressionoffers an important feedback to the mucin gene cluster to further retainnormal mucin and tissue homeostasis needed for optimal LRT function.

Membrane-tethered mucins line all wet surfaces of the body, but those ofthe LRT have the unique characteristic of fluctuating during a woman'shormonal cycle to allow at different times support for outflow of mensesblood from the uterus, the rapid transport of sperm cells into theFallopian Tube during ovulation, and the passage of a baby and placentaat child birth, all while stopping highly pathogenic bacteria fromascending to infect the host.

LRT mucins do much more than just hydrate the vaginal canal (Govindarian& Gipson, Exp. Eye Res. 90:655, 2010). Mucins in the vagina define azone between the epithelial cell layer and the external environment (thevaginal lumen). They create a 40-100 nm thick carbohydrate coating thatis anchored to the epithelium by proteins. This mucous layer excludesorganisms that are larger than 100 nm from directly contacting andinfecting the vaginal mucosal cell surface. For comparison, sperm areabout 50 μm in size. But, these mucins have been difficult to studybecause they are large, and their functional biochemistry is inseparablefrom their physical biochemistry, which is interactive with all thecells that make up the vaginal ecosystem (Kesimer & Sheehan, Glycobiol.18:463, 2008).

In the LRT, carbohydrate-rich mucus is secreted by vaginal epithelialcells and by cervical cells into the vaginal canal. This cervical mucus(CM) contributes to homeostasis and physiologic functioning of a woman'sLRT, which is a complex matrix of GAGs, mucins and other factors, suchas proteins and enzymes (Curlin and Bursac, Front. Biosci. 5:507, 2013).CM mucins are packaged in secretory granules and released, where theyswell to become a hydrogel in the vaginal canal (Espinosa et al., Hum.Reprod. 17:1964, 2002). Mucin hydration and gel formation are tightlycontrolled in the vagina and cervix by changes in calcium and pH levels.For example, an increase of calcium levels in the vaginal lumen from 1mM to 4 mM or a reduction in pH from 7.4 to 6 can decrease the speed atwhich CM mucin granules are hydrated and spread through the vagina,thereby negatively impacting sperm transport.

Overall, CM is composed of about 82% mucins, 3% HA, 2% heparin sulfateand 13% dermatan sulfate (Uldbjerg et al., 1983, Gynecol. Obstet.Invest. 16:199-209). Andersch-Bjorkman et al. (2007, Mol. CellProteomics 6:708) teach that altered mucus properties are correlatedwith fertility failure. The major mucin of CM is MUC5B, which peaks atovulation and is involved in sperm transport. Three gel forming mucinsare present in CM including MUC5B, MUC5AC, MUC6; and two transmembranemucins are present, including MUC1 and MUC16 (which is not foundanywhere else in the female reproductive tract). CM interestinglycontains several enzymes (such as amylase) also present in saliva.

For most of a woman's cycle, CM is scant, dry, sticky and thick, whichacts to block sperm and bacteria from ascending up the reproductivetract. But during ovulation, when the woman's estrogen levels rise, theCM can show a marked increase in water content and volume (up to20-fold). The fertile CM develops an “egg-white” quality, as it becomesmore slippery with a lower viscosity. Fibers in the fertile CM line upto form passageways for the sperm to rapidly penetrate through thecervix to reach the Fallopian tube, where the egg is waiting to befertilized (Chretien, Acta Obstet. Gynecol. Scand. 82:449, 2003). Duringovulation, increases in circulating estrogen increase water flow thruthe vaginal lining cells. This increased water in the vaginal canalhydrates secreted CM mucin granules to form the egg-white “fertile” CM.During hormone changes at ovulation, the vaginal environment supportsfertile CM production through enhancing water flow to hydrate thesecreted CM granules, decreasing calcium levels and a lowering of pH tostabilize and protect sperm. At ovulation, secreted granules of thesemucins begin to swell within 10 seconds after release from theepithelial cell, and they are totally dispersed within 30 minutes.Increased calcium levels reduce this expansion by one order ofmagnitude. Lowering of pH from 7 to 6.5 also results in an overall20-fold reduction in mucin granule swelling. Therefore, loss inhomeostasis in the LRT and particularly in the vaginal environmentduring ovulation can easily lead to thicker, less viscous CM thatimpedes sperm movement and fertility. Abnormal or “hostile” cervicalmucus that blocks sperm penetration into the Fallopian Tube duringovulation is a common cause of female infertility, with no currenttreatment.

Sperm cells and semen can also be a part of a healthy LRT and vaginalecosystem. During ovulation, a woman's body optimizes this environmentto support selection of healthy sperm to participate in fertilization.Contact of sperm with cervical mucins causes a dose related increase insperm velocity and linearity (straight swimming) that helps them reachthe egg (Eriksen et al., Fertil. Steril. 70:350, 1998).

A multitude of other changes in CM occurs during ovulation, includingthe switch from production of acidic sugars to neutral ones, to allow pHelevation needed for sperm transport. Also Van der Linden et al. (1992,Fertil. Steril. 57:573-7) showed a pre-ovulatory decrease in fructoseand glucose in CM, which likely maximizes sperm function. It is knownthat the CM provides an active selection gradient for the highestquality sperm in the ejaculate. Sperm that penetrate into the CM tend tohave superior DNA quality (Bianchi et al., J. Assist. Reprod. Genet.21:97, 2004) and lower lipid content, which prevents over activeoxidation damage (Feki et al., Mol. Hum. Reprod. 10:137, 2004). Also,contrary to the image of sperm “battling through” the cervical canal,out of the 250 million sperm deposited in a normal human ejaculate, only200 sperm end up in the Fallopian Tube near the site of fertilization.These sperm may actually be rapidly transported by the woman'sreproductive tract through the cervix and out of the vaginalenvironment. Sperm transport out of the vagina is seen in as little as90 seconds and is primarily complete within 30 minutes afterintercourse. The concerted actions of the LRT to ensure selection of thebest sperm for reproduction highlight the importance of vaginalecosystem homeostasis for overall healthy reproductive function.

Human sperm becomes capable of fertilizing the egg following enzymechanges (e.g., tyrosine phosphorylation) in the sperm membrane. If spermare to participate in fertilization, this shift cannot occur beforesperm meet the egg in the Fallopian Tube, but this physiologic changewas recently observed for sperm bound to GAG hyaluronic acid (HA) (Satiet al., Reprod. Sci. 2:573, 2014). In fact, sperm activation is exactlythe same if sperm are bound to an egg or to HA. Normally HA is a smallcomponent of CM at ovulation (just 3%), but changes in CM GAGcomposition could lead to abnormal sperm changes. In certainembodiments, compositions of this disclosure can be vaginally appliedformulations that alter GAG composition in the LRT (optionally with anHA based lubricant) to promote or enhance LRT homeostasis in the contextof fertility.

A. Compositions

Methods and compositions to maintain, preserve, promote, improve orenhance LRT homeostasis and healthy ecosystem function during cosmetic,prophylactic, and therapeutic interventions for the vulva, vagina andectocervix are provided herein. Also provided herein are methods andcompositions to maintain, preserve, promote, improve, enhance, orincrease sperm, oocyte, or embryo function or survival (e.g., duringassisted reproduction procedures). In certain embodiments, compositionsof this disclosure comprise isotonic formulations at a pH ranging fromabout 3.0 to about 8.0 that contains xylose and a Salvia extract. Thesecompositions are for topical application to support life-stage specificaspects (e.g., reproductive, post-menopausal) of LRT physiology andfunction. Preferably, the xylose/Salvia extract compositions furthercomprise additional components, such as a viscosity-increasing agent, asurfactant, a buffering agent, an osmolality adjuster, a pH modifier, asolvent, a preservative (e.g., paraben-free), a humectant or anycombination thereof. The inclusion of one or more these additionalcomponents will be dictated by the particular use (e.g., vaginalfreshening, perineal massage, fertility, vaginal hydration).

The compositions and methods provided herein are designed to work inconcert with the LRT ecosystem in a safe and highly efficacious mannerto support and potentiate physiologic functions at different stages in awoman's life and promote LRT homeostasis. Furthermore, in certainembodiments, compositions of the present disclosure can also function topromote healthy immune system function without overstimulation. Forexample, without wishing to be bound to any particular theory ofmechanism of action, compositions of the present invention can preventor treat a condition or symptom(s) of inflammation by reducing theactivation of the innate and/or acquired immune system and production ofpro-inflammatory cytokines.

Specifically, the methods and compositions of this disclosure provideisotonic, biome-friendly compositions designed for topical applicationto support LRT function and promote LRT homeostasis. The pH of thecompositions of this disclosure will differ according to uses directedto different life-stage specific events (e.g., trying-to-conceive,pregnancy, menopause) or hormonal cycle of a woman (e.g., ovulating orluteal phase of the cycle).

In certain aspects, compositions of the present disclosure comprise anisotonic, biome-friendly composition containing xylose and a Salviaextract, wherein the composition is formulated for topical applicationand has a pH ranging from about 3.0 to about 8.0.

Xylose is an aldopentose type monosaccharide having a chemical formulaof C₅H₁₀O₅ that is a building block for the hemicellulose xylan and isalso the first saccharide added to serine of proteoglycans by O-linkedglycosylation. Xylose can be obtained from the xylan-rich portion ofhemicelluloses present in plant cell walls and fiber and is also called“wood sugar.” Xylose is commercially available in purified form from avariety of vendors (e.g., Sigma-Aldrich, St. Louis, Mo.). Xylose mayalso be extracted and purified from a variety of plant sources,including wood, straw, corn, coconut shell, sugar cane bagasse,raspberries, loganberries, blackberries, guava, pears, broccoli,spinach, eggplant, peas, green beans, okra, cabbage, aloe vera,echinacea, boswellia, psyllium seeds, jute stick, rice husk, birch,alfalfa, leaf wood, apple and citrus pulp, sugar beet, and other plantmaterial. Pectic substances, also referred to as pectins, are a complexmixture of polysaccharides characterized by a backbone of α(1→4) linkedgalacturonic acid units that are partially methyl-esterified (O'Neill etal., Meth. Plant Biochem. 2:415, 1990). All pectins contain someassociated neutral sugars, such as L-arabinose, D-galactose, L-rhamnose,D-xylose, and D-glucose. Chemical and enzymatic degradation of pectinsreveal long and regular uronic regions (smooth) and rhamnose-richregions (hairy) that have neutral side sugars as side chains. Pectinsare present in the primary cell walls of all seed-bearing plants and aremajor components of dicotyledons (e.g., citrus and legumes) andgymnosperms (e.g., Douglas fir). Commercially important sources ofpectin include apple and citrus pulp, sugar beet, and alfalfa. Methodsof extracting and purifying xylose from natural sources are known in theart (see, e.g., Griffin et al., J. Chem. Tech. Biotech. 79:505, 2004;Ahmed et al., J. Biol. Sci. 1:1001, 2001; Saska and Ozer, Biotechnol.Bioeng. 45:517, 1995; and U.S. Pat. No. 3,687,807). D-xylose, which ispresent in living things, or L-xylose, which can be synthesized, can beused in the compositions disclosed herein. In certain embodiments,compositions of the instant disclosure comprise xylose having at least90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% purity. In someembodiments, xylose is from raspberries.

As discussed herein, carbohydrate GAGs are attached to the protein coreof proteoglycans (PGs) through the O-linked xylose sugar residue. Incertain embodiments, compositions of this disclosure containing xyloseare used to promote, maintain or achieve homeostasis of the vaginalmucosa or the lower reproductive tract. Without wishing to be bound bytheory, xylose may stimulate PG and/or GAG synthesis and secretion byvaginal tissues in ways that synchronize with hormonal and life cyclechanges of women, or in ways that protect the mucosal ecosystem of theLRT.

In certain embodiments, compositions of the instant disclosure comprisexylose at a concentration ranging from about 0.001% to about 10% byweight, about 0.001% to about 5% by weight, about 0.001% to about 2.5%by weight, about 0.001% to about 1% by weight, about 0.001% to about0.5% by weight, about 0.01% to about 5% by weight, about 0.01% to about2.5% by weight, about 0.01% to about 1% by weight, about 0.01% to about0.5% by weight, about 0.02% to about 1% by weight, or about 0.02% toabout 0.5% by weight. In certain embodiments, the concentration ofxylose is at about 0.001%, 0.005%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%,0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%,0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% by weight. Incertain embodiments, the xylose is D-xylose.

Compositions of the present disclosure also comprise a Salvia extract.Without wishing to be bound by theory, Salvia extracts may function bypreserving, improving or enhancing LRT homeostasis and healthy function.By way of background, Salvia (common name sage) is the largest genus ofplants in the mint family Lamiaceae, which includes almost 1,000species. The classification of Salvia is based on the genus' unusualpollination and stamen structure. Salvia species include annual,biennial, or perennial herbs, along with woody subshrubs. Severalmembers of the Salvia species have been used for their culinary,aromatherapy and medicinal value (see, e.g., Akhondzadeh et al., J.Clin. Pharm. Ther. 28: 53, 2003). There are three main geographicalregions of diversity, including Central and South America, Central Asiaand Mediterranean, and Eastern Asia.

Exemplary Salvia species include Salvia argentea, Salvia arizonica,Salvia azurea, Salvia candelabrum, Salvia carnosa, Salvia clevelandii,Salvia coccinea, Salvia columbariae, Salvia divinorum, Salvia dorrii,Salvia farinacea, Salvia forreri, Salvia fulgens, Salvia funerea, Salviaglutinosa, Salvia greggii, Salvia guaranitica, Salvia hispanica, Salvialeucantha, Salvia leucophylla, Salvia libanotica, Salvia longistyla,Salvia lyrata, Salvia mexicana, Salvia miltiorrhiza, Salvia patens,Salvia plebeia, Salvia polystachya, Salvia potus, Salvia pratensis,Salvia roemeriana, Salvia sclarea, Salvia spathacea, Salvia splendens,Salvia verticillata, and Salvia viridis (seewww.theplantlist.org/1.1/browse/A/Lamiaceae/Salvia for a complete listof Salvia species). In certain embodiments, Salvia extracts and oils canbe used in formulations of this disclosure to promote, improve, maximizeor maintain healthy function of the lower female reproductive tract.

In certain embodiments, a Salvia extract is a carbohydrate or oilextract from a Salvia species, such as an extract from Salvia plebeia,Salvia sclarea, Salvia hispanica, or any combination thereof.

Salvia plebeia (SP), also known as Asian sage, has been shown to have ananti-inflammatory and antioxidant activity (He et al., J. Cardiovasc.Pharmacol. 67:93, 2016), as well as anti-allergenic activity (Shi & Kim,Immunopharmacol. Immunotoxicol. 24:303, 2002). Salvia plebeia extractshave been shown to reduce bacterial LPS-activated inflammation andcytokine response (Akram et al., J. Ethnopharmacol. 174:322, 2015).Several different antioxidant and or anti-inflammatory active compoundshave been isolated from SP (Jin et al., Nat. Prod. Res. 29:1315, 2015;Akram et al., 2015) including, hispidulin, homoplantaginin, nepetin (twokinds), luteolin (two kinds), caffeic acid, luteoloside, eupatorin,hispidulin and plebeia oil A. These compounds show reactive oxygenspecies (ROS) scavenging at around 20 as well as production againstLPS-induced inflammation and COX-II inhibition, (Akram et al., 2015;Zhang et al., Molecules 20:14879, 2015).

Salvia sclarea (SS) extract, also known as Clary Sage, has been used asan aromatic essential oil for alleviating symptoms of depression andpremenstrual cramps. In fact, 5% clary sage has recently been shown tohave the strongest anti-depressant activity of any essential oil tested.This effect is due to its action on dopamine release (Seol et al., J.Ethnopharmacol. 130:187, 2010). Salvia sclarea extract has also beenshown to preserve cell viability and act as an antioxidant (Tavakkoli etal., 2014, Pharm Biol. 52:1550-7). The product sclaerol, a compoundfound in Salvia sclarea, inhibited the ability of interleukin inducedchondrocytes to trigger inflammatory pathways in vitro (Zhong et al.,In'tl. J. Clin. Exp. Pathol. 8:2365, 2015).

In addition, Salvia sclarea oil has been shown to have antimicrobialactivity, including acting synergistically with antibiotics to killresistant Staphylococcus organisms (Chovanova et al., Lett. Appl.Microbiol. 61:58, 2015). Salvia sclarea has also been shown to beeffective at killing common skin pathogens with little subsequentcytotoxicity (Sienkiewicz et al., Postepy Dermatol. Alergol. 32:21,2015; Firuzi et al., Iran J. Pharm. Res. 12:801, 2013; Cui et al.,Botanical Studies 56:1, 2015).

The International Federation of Professional Aromatherapists recentlyissued a Pregnancy Guidelines (2013)(www.naha.org/assets/uploads/PregnancyGuidelines-Oct11.pdf) foraromatherapists working with pregnant patients. Salvia sclarea is listedas an essential oil that may be used during pregnancy.

In certain embodiments, the total concentration of Salvia extract incompositions of this disclosure ranges from about 0.001% to about 30% byweight, about 0.001% to about 10% by weight, about 0.001% to about 5% byweight, 0.001% to about 1% by weight, about 0.001% to about 0.5% byweight, about 0.01% to about 5% by weight, about 0.01% to about 2.5% byweight, about 0.01% to about 1% by weight, about 0.01% to about 0.5% byweight, about 0.025% to about 1% by weight, about 0.025% to about 0.5%by weight, or about 0.01% to about 0.2% by weight. In furtherembodiments, the total concentration of Salvia extract is at about0.001%, 0.005%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%,0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%,4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%,19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, or 30% by weight.

In particular embodiments, compositions of the instant disclosurecomprise: (a) xylose at a concentration ranging from about 0.001% toabout 10% by weight, and Salvia extract at a concentration ranging fromabout 0.001% to about 30% by weight; (b) xylose at a concentrationranging from about 0.01% to about 5% by weight, and Salvia extract at aconcentration ranging from about 0.001% to about 30% by weight; (c)xylose at a concentration ranging from about 0.02% to about 1% byweight, and Salvia extract at a concentration ranging from about 0.001%to about 30% by weight; (d) xylose at a concentration ranging from about0.001% to about 10% by weight, and Salvia extract at a concentrationranging from about 0.01% to about 5% by weight; (e) xylose at aconcentration ranging from about 0.01% to about 5% by weight, and Salviaextract at a concentration ranging from about 0.01% to about 5% byweight; (f) xylose at a concentration ranging from about 0.02% to about1% by weight, and Salvia extract at a concentration ranging from about0.01% to about 5% by weight; (g) xylose at a concentration ranging fromabout 0.001% to about 10% by weight, and Salvia extract at aconcentration ranging from about 0.01% to about 1% by weight; (h) xyloseat a concentration ranging from about 0.01% to about 5% by weight, andSalvia extract at a concentration ranging from about 0.01% to about 1%by weight; or (i) xylose at a concentration ranging from about 0.02% toabout 1% by weight, and Salvia extract at a concentration ranging fromabout 0.01% to about 1% by weight.

The pH of the compositions disclosed herein range from about 3.0 toabout 8.0, depending on the stage of hormone cycle and/or stage of lifeof the female subject as described herein. In certain embodiments, thefemale subject is human.

The compositions of the present disclosure may further comprise anon-irritating viscosity increasing agent. Viscosity is a property ofliquids that is closely related to the resistance to flow. It may bedefined by Couette flow, which is the laminar flow of a viscous fluid inthe space between two parallel plates, one of which is moving relativeto the other. The flow is driven by virtue of viscous drag force actingon the fluid and the applied pressure gradient parallel to the plates.

In certain embodiments, compositions of the instant disclosure areformulated to have rheological properties best suited for the targettissue (i.e., female lower reproductive tract) and to mimic theproperties of normally occurring fluids and mucus surrounding exposedLRT cells. For example, compositions formulated as gels applied tomucous membranes may be designed to have viscosity values consistentwith or similar to normal mucus, and exhibiting non-Newtonian,shear-thinning (pseudoplastic) flow properties.

Standardized methodology for quantitative comparisons ofover-the-counter vaginal products based features such as, stickiness,ropiness, peaking, rubberiness, thickness, smoothness, and slipperiness,are known in the art (Mahan et al., Contraception, 2011, 84:184-193).

For compositions applied to skin (such as the vulva) or inside thevagina, a non-irritating viscosity increasing agent can be added in anamount that allows the composition to spread easily to form a thin layerwhen minimal physical pressure is applied, and to have adequateviscosity and shear-thinning properties so that the composition does not“run” off or out of the lower reproductive tract tissue upon topicalapplication. Mucoadhesive formulations that are retained at the vaginalmucosal surface for prolonged biological activity are known in the art(reviewed by Khutoryanskiy, Macromol. Biosci. 11:748, 2011; Brooks,Front. Chem. 3:65, 2015)

The rheological characteristics of compositions once present on thetissue of interest are complex and can be influenced by a variety offactors, including the formulation of the composition, temperature, pH,salt concentration, and ion concentration, as well as the presence andphysical properties of the physiological secretions (e.g., mucus, semen,blood, etc.).

In any of the aforementioned embodiments, the composition furthercomprises a non-irritating viscosity increasing agent at a concentrationof about 0.001% to about 45%, about 0.001% to about 30%, about 0.001% toabout 25%, about 0.001% to about 20%, about 0.001% to about 15%, about0.001% to about 10%, about 0.001% to about 5%, about 0.001% to about 3%,about 0.01% to about 30%, about 0.01% to about 25%, about 0.01% to about20%, about 0.01% to about 15%, about 0.01% to about 10%, about 0.01% toabout 5%, about 0.01% to about 3%, about 0.1% to about 30%, about 0.1%to about 25%, about 0.1% to about 20%, about 0.1% to about 20%, about0.1% to about 15%, about 0.1% to about 10%, about 0.1% to about 5%,about 0.1% to about 3%, about 0.5% to about 30%, about 0.5% to about25%, about 0.5% to about 20%, about 0.5% to about 15%, about 0.5% toabout 10%, about 0.5% to about 5%, about 0.5% to about 3%, about 1% toabout 30%, about 1% to about 25%, about 1% to about 20% about 1% toabout 15%, about 1% to about 10%, about 1% to about 5%, about 1% toabout 3%, about 5% to about 30%, about 5% to about 25%, about 5% toabout 20%, about 5% to about 15%, or about 5% to about 10% by weight.

Non-irritating viscosity-increasing agents for use in the compositionsof the present disclosure are used in concentrations that do not causecell or vaginal microbiota toxicity, sperm toxicity, oocyte toxicity,embryo toxicity, epithelial cell damage, cytokine release, or mucusfunction disruption. In certain embodiments, a non-irritatingviscosity-increasing agent is a carbomer, polyoxazoline, cellulose,cellulose ether, or a combination thereof. A cellulose ether may bemethylcellulose, ethylcellulose, ethyl methyl cellulose, hydroxyethylcellulose, hydroxypropyl cellulose, hydroxyethyl methyl cellulose,hyroxypropyl methyl cellulose (hypromellose), ethyl hydroxyethylcellulose, carboxymethyl cellulose, or any combination thereof. Inparticular embodiments, a non-irritating viscosity-increasing agent isselected from cetyl hydroxyethylcellulose, carbomer homopolymer type A,carbomer homopolymer type B, carbomer homopolymer type C, hydroxypropylmethyl cellulose (HPMC or hypromellose), carbomerhomopolymer/hypromellose, hydroxyethyl cellulose,hydrophobically-modified hydroxyethyl cellulose, hydroxypropylcellulose, carboxymethyl cellulose, ethylcellulose, ethyl methylcellulose, hydroxyethyl methyl cellulose, hydroxyethyl methyl cellulose,ethyl hydroxyethyl cellulose, methylcellulose, guar gum, polyacrylate,acrylate copolymer, acrylates crosspolymer-4, acrylates/C₁₀₋₃₀ alkylacrylate crosspolymer, carbomer interpolymer Type A, carbomerinterpolymer Type B, carbomer copolymer Type A, carbomer copolymer TypeB, polycarbophil, polyvinyl alcohol, polyvinylpyrrolidone,polyoxazoline, or any combination thereof.

In further embodiments, polyoxazoline is a poly(2-oxazoline), forexample, a poly(2-alkyl-2-oxazoline) or poly(2-aryl-2-oxazoline). Insome embodiments, the poly(2-alkyl-2-oxazoline) ispoly(2-methyl-2-oxazoline), poly(2-ethyl-2-oxazoline),poly(2-isopropyl-2-oxazoline), or any combination thereof. In particularembodiments, the poly(2-aryl-2-oxazoline) is poly(2-phenyl-2-oxazoline).Polyoxazolines, such as poly(2-ethyl-2-oxazoline) (PetOx) and relatedchemicals, offer high levels of biocompatibility, as shown in mucosalirritation studies. These polymers do not decompose under physiologicalconditions, so no oxidative triggering products are produced (VanKuringen et al., Macromol. Biosci. 12:1114, 2012).

In certain embodiments, a non-irritating viscosity-increasing agentcomprises cellulose ether and a carbomer. In some embodiments, thecellulose ether is methylcellulose, ethylcellulose, ethyl methylcellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxyethylmethyl cellulose, hyroxypropyl methyl cellulose (hypromellose), ethylhydroxyethyl cellulose, carboxymethyl cellulose, or any combinationthereof. In further embodiments where hydration of vaginal mucosa andsecretions are desired, the ratio of cellulose ether (e.g.,hypromellose) to carbomer in the composition ranges from about 1:1 toabout 8:1, from about 2:1 to about 8:1, or from about 2.5:1 to about6.5:1. In further embodiments where muco-adherence and product retentionin the vagina are desired, the ratio of cellulose ether (e.g.,hypromellose) to carbomer in the composition is from about 1:1 to 1:5 orfrom about 1:2 to about 1:4. In a specific embodiment, the ratio ofcellulose ether (e.g., hypromellose) to carbomer in the composition isabout 1:2, 2.6:1, 3.3:1, or about 6.2:1.

The tissue muco-adhesion properties, and consequently residence-time onthe LRT tissue, of the compositions disclosed herein may vary dependingon its intended use. For example, if a composition is formulated toadhere to the vaginal mucosa for several hours, for 24 hours, or fordays, a suitable non-irritating viscosity-increasing agent may be usedthat allows for muco-adhesion for that period of time.

In some embodiments, compositions of the present disclosure have athicker viscosity for muco-adhesion and balancing of pH, or delivery ofdrugs over a longer time frame. In other embodiments, compositions ofthe present disclosure have a lower viscosity for maximal function. Forexample, lower viscosity compositions provide increased comfort forhydrating formulations, decreased friction during sexual intercourse forlubricating formulations, or rehydration of vaginal secretions andcervical mucus granules for fertility enhancing formulations.

Methods of measuring viscosity of non-Newtonian fluids are known in theart and include rotational rheometry techniques. Other techniques formeasuring viscosity are the rolling ball viscometer method (Hubbard andBrown, Ind. Eng. Chem. Anal. Ed. 15: 212-218, 1943) and the capillaryviscometry method.

In certain embodiments, the viscosity of the compositions of the presentdisclosure is about 5 centipoise (cps) to about 750,000 cps, about 500cps to about 600,000 cps, about 1,500 cps to about 600,000 cps, about5,000 to about 500,000 cps, or about 10,000 to about 100,000. In someembodiments, the viscosity of the compositions of the present disclosureis about 50 cps to about 5,000 cps, about 1,000 cps to about 15,000 cps,about 10,000 cps to about 75,000 cps, about 50,000 cps to about 200,000cps, or about 100,000 cps to about 600,000 cps.

Compositions of the present disclosure may further comprise a solvent. Asolvent may be an aqueous solvent, a hydrophobic solvent, or acombination thereof. An aqueous solvent may be water. The hydrophobicsolvent may be pharmaceutical grade mineral oil or pure, non-oxidizedoils made from the plants of: rose, raspberry, apricot, jojoba, olive,corn, cottonseed, peanut, soybean, sesame, or a combination thereof. Ifoils are used as a solvent in the present composition, it is desirablethat the oils are non-oxidized, as many oils have oxidation products,including peroxide, that can cause irritation and immune stimulation inthe female LRT. In some embodiments, the solvent is water. The solventmay be at a concentration ranging from about 50% to about 98%, about 60%to about 98%, about 65% to about 98%, about 70% to about 98%, about 75%to about 98%, about 80% to about 98%, about 85% to about 98%, about 90%to about 98%, or about 95% to about 98% by weight in the composition.

In certain embodiments, compositions of the present disclosure mayfurther comprise a buffering agent. A buffering agent contains an acidicspecies to neutralize hydroxide (OH) ions and a basic species toneutralize hydrogen (H⁺) ions. However, the acidic and basic species ofthe buffering agent should not consume each other through aneutralization reaction. The buffering agent can be a weak acid and asalt of the weak acid or a weak base and a salt of the weak base. Thus,the buffering agent can include a weak acid-base conjugate pair or weakbase-acid conjugate pair. In certain embodiments, the buffering agent isselected such that the buffering agent's acid form has a pKa the same asor close to the desired pH of the composition or a pH within the desiredrange of pH values. The total amount of the buffering agent (e.g.,conjugate acid-base pair) is selected such that the pH of thecomposition is maintained at the desired pH or range of pH values. Thebuffering capacity is the amount of acid or base the buffer canneutralize before the pH of the composition begins to change to anappreciable degree. Thus, the greater the amount of the buffering agent,the more resistant the pH of the composition is to change. In certainembodiments, a buffering agent comprises a monocarboxylate, adicarboxylate, a carboxylic acid, or a combination thereof. In someembodiments, a buffering agent may comprise an acetate, borate, citrate,fumarate, lactate, malate, malonate, nitrate, phosphate, propanoate,succinate, tartrate, tromethamine, or any combination thereof. In someembodiments, a buffering agent comprises lactic acid, sodium lactate,sodium phosphate (monobasic, dibasic, or both), potassium phosphate(monobasic, dibasic, or both), sodium citrate, potassium citrate, aceticacid, sodium acetate, citric acid, disodium citrate, trisodium citrate,boric acid/sodium, succinic acid, sodium succinate, disodium succinate,tartaric acid, sodium tartarate, sodium ascorbate, ascorbic acid,tromethamine (Tris), or any combination thereof. In further embodiments,a buffering agent comprises sodium lactate and lactic acid, sodiumphosphate and potassium phosphate, sodium phosphate and potassiumcitrate, sodium phosphate, sodium citrate, or ascorbic acid and sodiumascorbate.

In certain embodiments, a buffering agent may be at a concentration fromabout 0.05% to about 4% by weight, about 0.1% to about 3% by weight,0.25% to about 2% by weight, or about 0.5% to about 1% by weight in thecomposition. For those buffering agent acids that comprise monobasic,dibasic, or tribasic forms (e.g., potassium phosphate, sodiumphosphate), as used herein, general reference to the acid includes themonobasic, dibasic, and tribasic forms. The amount of the acid means thetotal amount of any monobasic, dibasic, and tribasic acid forms present.For example, a composition comprising 1% sodium phosphate by weight maycomprise monobasic sodium phosphate and dibasic sodium phosphatetotaling 1%.

Topical compositions of the present disclosure are formulated at a pHequivalent to the healthy physiologic pH for the LRT tissue to which itis administered and lifecycle stage of the female subject, or for thetherapeutic goals of the composition. The buffer agent selected willhave a buffering capacity adequate to maintain pH within the healthyphysiologic range for the target LRT tissue during its time of residenceon the tissue, or to achieve the therapeutic goals of the composition.In certain embodiments, the buffer capacity of the composition issufficient to resist adjustment by fluid and to maintain the target pHfor an appreciable period of time.

In certain embodiments, a composition may simultaneously come intocontact with two tissues or cell types that differ in pH (e.g., vaginalmucosa and sperm cells). Special care in formulating may be needed inthese circumstances regarding pH and buffer capacity. Some tissues orcells can tolerate less deviation from physiological conditions comparedto others (e.g., sperm cells versus vaginal mucosal cells).

In certain embodiments, a composition of this disclosure may furthercontain an active ingredient (e.g., drug). The pH of the drug containingcomposition may be adjusted to maintain stability of the activeingredient. In certain embodiments, the buffering capacity of acomposition is adequate for drug stability and also matches the overallpH of the target tissue or therapeutic goals of the composition.

In certain embodiments, compositions of the present disclosure furthercomprise a pH modifying agent. The pH modifying agent may comprise anacidifying agent, an alkalinizing agent, or both. In some embodiments,the acidifying agent is acetic acid, citric acid, fumaric acid,hydrochloric acid, lactic acid, malic acid, nitric acid, phosphoricacid, potassium phosphate monobasic, propionic acid, sodiumphosphatemonobasic, sulfuric acid, tartaric acid, or any combinationthereof. In some embodiments, the alkalinizing agent is ammonia,ammonium carbonate, diethanolamine, monoethanolamine, potassiumhydroxide, potassium phosphate dibasic, sodium bicarbonate, sodiumborate, sodium carbonate, sodium hydroxide, sodium lactate, sodiumphosphate dibasic, trolamine, or any combination thereof.

In certain embodiments, a pH modifying agent may be at a concentrationranging from about 0.001% to about 2%, from about 0.005% to about 0.75%,from about 0.01% to about 1%, from about 0.01% to about 0.5%, or fromabout 0.02% to about 0.4% by weight of the composition.

The pH range of the compositions of the present disclosure is from about3.0 to about 8.0, depending on the physiologic stage of the woman andthe intended use of the product. In certain embodiments, the pH range ofthe composition is about 3.5 to about 5.5 (e.g., for a reproductive agedfemale), about 4.5 to about 6.8 (e.g., for a menopausal female), about4.5 to about 6.5 (e.g., for a pregnant female), or about 6.8 to about7.8 (e.g., for a reproductive aged female during ovulatory phase). Incertain embodiments, the pH range of a composition may be broad enoughto be targeted to female subjects in different life/cycle stages. Forexample, a composition may have a pH range of about 3.5 to about 6.8 andbe used in a reproductive aged, non-ovulating female subject, amenopausal female subject, and a pregnant female subject.

In certain embodiments, compositions of the present disclosure aresterile and preservative-free. In other embodiments, sterile,preservative-free compositions are in a single-use or unit-dose format.

In other embodiments, compositions of the present disclosure optionallycomprise a preservative. In particular embodiments, a preservative is aparaben-free preservative. Parabens are a series of parahydroxybenzoatesor esters of parahydroxybenzoic acid and are known to cause cytokinerelease and irritation and have been linked to several types of cancer.Examples of parabens include methylparaben, ethylparaben, propylparaben,butylparaben, heptylparaben, isobutylparaben, isopropylparaben,benzylparaben, and their sodium salts.

Exemplary paraben-free preservatives include phenethyl alcohol, caprylylglycol, phenoxyethanol, a sorbate, potassium sorbate, sodium sorbate,sorbic acid, sodium benzoate, benzoic acid, acemannan, oleuropein,carvacrol, cranberry extract, gluconolactone, green tea extract,Helianthus annuus seed oil, Lactobacillus ferment, Usnea barbataextract, polyaminopropyl biguanide, polyglyceryl-3 palmitate,polyglyceryl-6 caprylate, pomegranate extract, Populus tremuloides barkextract, resveratrol, Rosmarinus officinalis leaf extract, benzylalcohol, or any combination thereof. In some embodiments, compositionsof the present disclosure include a preservative in combination with oneor more paraben-free preservative, such as phenethyl alcohol andcaprylyl glycol. In particular embodiments, a paraben-free preservativecomprises phenethyl alcohol in an amount of 56%-60% and caprylyl glycolin an amount of 40%-44% (e.g., Tristat Stabil).

In certain embodiments, compositions of this disclosure further comprisea preservative at a concentration of about 0.005% to about 10% byweight, about 0.005% to about 5%, about 0.005% to about 2.5%, about0.005% to about 1%, about 0.005% to about 0.5%, about 0.005% to about0.1%, about 0.01% to about 10%, about 0.01% to about 5%, about 0.01% toabout 2.5%, about 0.01% to about 1%, about 0.01% to about 0.5%, about0.01% to about 0.1%, about 0.1% to about to about 10%, about 0.1% toabout 5%, about 0.1% to about 2.5%, about 0.1% to about 1%, about 0.1%to about 0.5%, about 1% to about 10%, about 1% to about 5%, or about 1%to about 2.5% by weight.

In further embodiments, compositions of the present disclosure furthercomprise an osmolality adjuster or osmolality adjusting agent.Osmolality is a measure of minimum pressure which needs to be applied toa solution to prevent the inward flow of water across a semipermeablemembrane and is expressed in milliosmoles per kilogram of solvent(mOsmol/kg). The osmotic pressure of a solution depends on the number ofparticles in solution. Osmolality of a solution may be measured bymeasuring freezing point depression or vapor pressure lowering.

In certain embodiments, the compositions disclosed herein are bothiso-osmotic and isotonic with respect to the targeted lower reproductivetract tissues and the mucus they secrete. Compositions havingphysiologic osmolality maintain LRT cell structure and function.Exposure of LRT cells to high osmolality compositions will cause a netmovement of water out of the cells and irreversible ultrastructuraldamage. Exposure of LRT tract cells to low osmolality compositions willcause a net movement of water into the cells and may lead to cellbursting/death. Iso-osmotic compositions, having the same osmolality asthat within LRT cells or LRT mucus secretions, are safe for use in theLRT.

Tonicity is a measure of the effective osmotic pressure gradient (asdefined by the water potential of two solutions) of two solutionsseparated by a semipermeable membrane. Tonicity is commonly used whendescribing the response of cells immersed in an external solution. Inother words, tonicity is the relative concentration of solutions thatdetermine the direction and extent of diffusion. Body fluids normallyhave an osmotic pressure that corresponds to that of a 0.9% solution ofsodium chloride. A composition (e.g., solution or gel) is consideredisotonic when its tonicity is about equal to that of a 0.9% sodiumchloride solution (i.e., 290 mOsm). A composition is isotonic with abody fluid solution when the magnitude of the salts are equal betweenthe composition and the physiologic solution. Tonicity equilibrium isreached in physiologic solutions by water moving across the membranes,but the salts staying in their solution of origin. A solution isisotonic with a living cell if there is no net gain or loss of water bythe cell, or other changes in the cell, when it is in contact with thatsolution.

Hypertonic solutions cause a net movement of water out of the cells (asthe water moves to create equilibrium with the high salt levels outsideof the cell). This dehydration of the cell is concentration dependentand leads to osmotic stress which can increase reactive oxygen species,cause cytoskeletal rearrangement, and damage DNA and mitochondrialfunction within minutes of exposure. Hypotonic solutions cause a netflow of water into the cell and cause cell bursting and death. In orderto maximize homeostasis and healthy function of sperm and lowerreproductive tract mucosal cells, the compositions of the presentdisclosure are isotonic.

The term isotonic, meaning equal tone, is commonly used interchangeablywith the term iso-osmotic. However, iso-osmotic actually is a more broadterm that compares the osmotic pressure of two liquids, wherein thesmall particles present need not be salts. The particles providingosmotic pressure can be membrane permeable compounds such as glycerol orpropylene glycol. In this case, water and the osmotic particles both canmove across the membranes. The osmotic particles do not move in responseto highly orchestrated cell transport concentration gradients butprimarily through diffusion. Therefore, these particles end up insideliving cells during physiologic exposure. Often such particles causedamage inside the cell they penetrate. For example, a solutioncontaining glycerol that is iso-osmotic with semen, has the sameosmolarity as semen, but the glycerol can penetrate into the sperm celland damage its function. Osmolarity takes into account the totalconcentration of penetrating solutes and non-penetrating solutes,whereas tonicity takes into account the total concentration of onlynon-penetrating solutes. Isotonicity infers a sense of physiologicalcompatibility whereas iso-osmotic does not require this.

In certain embodiments, an osmolality adjuster used in the compositionsdisclosed herein is an electrolyte, mono- or disaccharide, inorganicsalt (e.g., sodium chloride, calcium chloride, sodium sulfate, magnesiumchloride), or a combination thereof. In some embodiments, an osmolalityadjuster is glucose, sucrose, sodium chloride, potassium chloride,calcium chloride, sodium sulfate, magnesium chloride, dextrose,mannitol, or any combination thereof.

In certain embodiments, the osmolality range of the compositionsdisclosed herein is about 200 mOsm/kg to about 600 mOsm/kg, about 240mOsm/kg to about 400 mOsm/kg, or about 260 mOsm/kg to about 380 mOsm/kg.

In certain embodiments, compositions disclosed herein further comprise asurfactant. In some embodiments, the surfactant is cetylhydroxyethylcellulose, hydrophobically modified hydroxyethyl cellulose,poloxamer, polyoxyethylene glycol alkyl ether, polyoxypropylene glycolalkyl ether, glucoside alkyl ether, polyoxyethylene glycol alkylphenolether, glycerol alkyl ester, polysorbate, cocamide monoethaolamine(MEA), cocamide diethanolamine (DEA), dodecyldimethylamine oxide, or anycombination thereof. The surfactant may be present at a concentration ofabout 0.01% to about 20%, about 0.01% to about 15%, about 0.01% to about10%, about 0.01% to about 10%, about 0.01% to about 5%, about 0.01% toabout 2.5%, about 0.01% to about 1%, about 0.01% to about 0.5%, about0.01% to about 0.1%, about 0.1% to about 20%, about 0.1% to about 10%,about 0.1% to about 5%, about 0.1% to about 2.5%, 0.1% to about 1%, orabout 0.1% to about 0.7%, about 1% to about 20%, about 1% to about 10%,about 1% to about 15%, about 1% to about 10%, about 1% to about 5%, orabout 1% to about 2.5% by weight.

In certain embodiments, the compositions disclosed herein possess anendotoxin level of less than or equal to 20 EU/ml, 15 EU/ml, 10 EU/ml,7.5 EU/ml, 5 EU/ml, 2.5 EU/ml, 1 EU/ml, 0.75 EU/ml, 0.5 EU/ml, 0.25EU/ml, or 0.1 EU/ml. Bacterial endotoxins, found in the outer membraneof gram-negative bacteria, are members of a class of phospholipidscalled lipopolysaccharides (LPS). LPS and endotoxins are powerful innateimmune system stimulants. Even picogram concentrations of LPS canstimulate chronic inflammatory conditions leading to systemic andlocalized disease.

Bacterial endotoxin levels can be quantified using methods known in theart, including the Limulus Amebocyte Lysate (LAL) assays. This assay isbased in the biology of the horseshoe crab (Limulous). These animalsproduce LAL enzymes in blood cells (amoebocytes) to bind and inactivateendotoxin from invading bacteria. The inactivation of endotoxin by LALforms a clot. The LAL test exploits the clotting action of this enzyme,by adding a LAL reagent to the tested product, and assaying for clotformation. LAL test methodologies include gel-clot techniques,photometric techniques including kinetic turbidimetric methods, andchromogenic methods.

Compositions of the present disclosure may optionally comprise ahumectant, an emollient, or both. In some embodiments, the humectant isa sugar alcohol, such as glycerin, sorbitol, xylitol, mannitol; hexyleneglycol, butylene glycol, propylene glycol glyceryl triacetate, an alphahydroxyl acid (e.g., lactic acid), galactoarabinan, or a combinationthereof. In some embodiments, the emollient is pharmaceutical grademineral oil, a purified, non-oxidized plant oil (e.g., rose, raspberry,corn, cottonseed, peanut, soybean, sesame, apricot, jojoba, olive),cetostearyl alcohol, glyceryl stearate, acemannan, or a combinationthereof.

The function of each of the excipients is not mutually exclusive withinthe context of the compositions of the present disclosure. For example,cetyl hydroxyethylcellulose or hydroxyethyl cellulose may be used as anon-irritating viscosity increasing agent or a surfactant. In yetanother example, sodium lactate or lactic acid may be used as abuffering agent or a pH modifier. In yet another example, acemannan maybe used as a paraben-free preservative, an emollient, or a prebiotic.

In certain embodiments, the compositions disclosed herein furthercomprise an additional therapeutic agent. For example, a therapeuticagent may be included in the composition to improve cell or tissuefunction or to treat an underlying disease or disorder. In anotherexample, a therapeutic agent may be an anti-microbial agent (e.g., anantibiotic, anti-fungal agent, anti-viral agent, or any combinationthereof). In certain embodiments, an anti-microbial agent is ananti-fungal agent selected from butoconazole nitrate, clotrimazole,miconazole nitrate, terconazole, tioconazole, econazole nitrate,efinaconazole, ketoconazole, luliconazole, naftifine hydrochloride,oxiconazole nitrate, sertaconazole nitrate, sulconazole nitrate,tavaborole, terbinafine, acyclovir, tenovir, zidovudine, stavudine,metronidazole, or a combination thereof.

Other examples of therapeutic agents include hormones (e.g., estradiol,estriol, estropipate, testosterone, progesterone, DHEA or a combinationthereof) for use in treating genitourinary syndrome of menopause (GSM),agents for treating or preventing a hormone imbalance or infertility(e.g., progesterone, estrogen, testosterone), contraceptives (e.g.,impairs sperm function, thickens cervical mucus, or both), growthfactors (e.g., vascular endothelial growth factor), vasodilators (e.g.,L-arginine, niacin, nicotinamide, alprostadil, phosphodiesteraseinhibitor).

In certain embodiments, the compositions disclosed herein furthercomprise factors that maintain, improve, or enhance VMB (e.g.,Lactobacilli) function and health, such as amylase, glycogen, D-lacticacid, L-lactic acid, or a combination thereof.

In certain embodiments, the compositions disclosed herein furthercomprise at least one prebiotic. A prebiotic refers to non-digestibleplant fiber that stimulates the growth or activity of one or more VMBspecies. Examples of prebiotics that can be used include acemannan andgalactoarabinan (see, Al-Ghazzewi and Tester, 2016, J. Appl. Microbiol.10.1111/jam.13054 (e-publication ahead of print); Gullon et al., 2015,Food Funct. 6:525-531;http://www.fda.gov/ucm/groups/fdagov-public/@fdagov-foods-gen/documents/document/ucm266729.pdf).

In certain embodiments, the compositions disclosed herein furthercomprise at least one vaginal probiotic bacterial species or strain(e.g., belonging to the genus Lactobacillus). As used herein, “vaginalprobiotic bacteria” refer to live bacteria, which when administered inadequate amounts to the vagina confer a health benefit to the host. A Incertain embodiments, the probiotic bacterial species or strain is onehaving the ability to colonize the human vagina. The adhesion oflactobacilli to the uroepithelium varies among species and strains, asshown by in vitro studies (Reid et al., 1987, J. Urol. 138:330-335), andmay be mediated by glycoprotein and carbohydrate adhesins binding toglycolipid receptors (Boris et al., 1998, Infection and Immunity66:1985-1989). In some embodiments, a vaginal probiotic species is aspecies that is part of the VMB. In a specific embodiment, a vaginalprobiotic species is selected from Lactobacillus fermentum,Lactobacillus acidophilus, Lactobacillus jensenii, Lactobacillusgasseri, Lactobacillus iners, Lactobacillus crispatus, Lactobacillusplantarum, Lactobacillus fermentum, Lactobacillus brevis, Lactobacilluscasei, Lactobacillus delbrueckii, Lactobacillus vaginalis, Lactobacillussalivarius, or any combination thereof. In some embodiments, a vaginalprobiotic bacterial species or strain is encapsulated or coated.Compositions containing a vaginal probiotic species preferably include aparaben-free preservative.

A vaginal probiotic bacterial species or strain may be administered inan amount of about 10⁴ to about 10¹² colony forming units (CFU), orabout 10⁵ to about 10¹¹ CFU, or about 10⁶ to about 10¹⁰ CFU, or about10⁷ to about 10⁹ CFU per dose. In some embodiments, a vaginal probioticbacterial species or strain may be administered in an amount of about 4million to about 400 million colony forming units (CFU) per dose.

A vaginal probiotic bacterial species or strain may be included in thecompositions disclosed herein for treatment and/or prophylaxis ofbacterial vaginosis, viral vaginosis, yeast vaginitis, infections in thevagina, sexually transmitted diseases, such as HIV and chlamydiainfection, infections endangering the fetus in pregnant women, pretermlabour and urinary tract infection.

In certain embodiments, the compositions of the present disclosure donot comprise polyethylene glycol (PEG). Because of its widespread use,antibodies to PEG are seen in 25% of patients. PEG is prone to oxidativedegradation that can further trigger an immune response in patients (Dela Rosa, Mater Sci. Mater Med. 25:1211, 2014). PEG triggers high levelsof interleukin secretions in cervical cells (Gali et al., Antimicrob.Agents Chemother. 54:5105, 2010).

In certain embodiments, the compositions of the present disclosure donot comprise a glycosaminoglycan (GAG), for example hyaluronic acid.Although GAGs naturally exist in the vaginal ecosystem, use of purifiedGAGs for treating vaginal dryness may cause a disruption in the LRThomeostasis and function (Chen et al., J. Sex. Med. 10:1575, 2013).Hyaluronic acid, a common ingredient in personal lubricants, has beenshown to cause vaginal epithelial cells to release changed antimicrobialpeptides via TLR signaling. Elevated hyaluronan levels were associatedwith vaginal itching and burning or itching and discharge in women withvulvovaginal candidiasis and may contribute to symptoms (Lev-Sagie etal., 2009, Am. J. Obstet. Gynecol. 201:206).

In certain embodiments, compositions of the present disclosure do notcomprise glycerol. Glycerol has a direct toxic effect on sperm cellsdamaging their cell membranes and metabolism as it penetrates the cell(McLaughin et al., 1992, J. Reprod. Fertil. 95:749-54). Glycerol alsocauses a two-fold increase in interleukin production from cervical cellsat a 0.5% concentration and a 5-fold increase in interleukin at a 5%concentration. This concentration range is common for many vaginalproducts on the consumer shelf.

In certain embodiments, compositions of the present disclosure do notcomprise an oil other than Salvia extract oil unless the oil ispharmaceutical grade or free from significant levels of oxidationproducts. An oil may be a petroleum oil, e.g., mineral oil, a plant oil(e.g., almond oil), or an essential oil. Photo-oxidation andauto-oxidation of oils occurs routinely. Oxidation changes in oils canhave a profound impact on biological function (Morbeck et al., 2010,Fertil. Steril. 94:2747-52). While peroxide levels of 10 meq/kg in oilsare considered safe for consumption, small concentrations (<1 meq/kg)causes reductions in in vitro sperm motility and embryo development. An8-fold increase in yeast infection rates has been attributed tonon-pharmaceutical grade mineral oil use in the vaginal ecosystem.Pharmaceutical grade mineral oil (“mineral oil USP”) is mineral oil thatbeen washed with pharmaceutical grade, highly purified water and testedfor peroxide value (POV) below 0.1 mEq.

One concern relating to the safety assessment of botanical ingredientsin personal care products is their contamination with heavy metals. Ifheavy metals are present in topically applied products, local andsystemic toxic and inflammatory effects may result. Heavy metals whenapplied to epithelial surfaces, such as the female lower reproductivetract, result in the generation of highly reactive oxygen free radicals,such as hydroxyl radical, thus stimulating oxidative damage to thetissues. Generation of highly reactive oxygen free radicals may alsoaccelerate degradation of the compositions of the present disclosure. Ifabsorbed systemically in sufficient quantities, some heavy metals mayhave toxic effects on cardiovascular, pulmonary, urinary,gastrointestinal, and neurological systems.

Heavy metals include cadmium, lead, inorganic arsenic, inorganicmercury, iridium, osmium, palladium, platinum, rhodium, ruthenium,chromium, molybdenum, nickel, vanadium, and copper.

The toxicity of an elemental impurity is related to its extent ofexposure (bioavailability). The extent of exposure has been determinedfor each of the elemental impurities of interest. These limits are basedon chronic exposure. Permissible daily exposure (PDE) described in Table1 below would apply to a composition of this disclosure to be applied tothe skin or mucosa.

The values provided in Table 1 represent concentration limits forcomponents (drug substances and excipients) of topical products appliedat a maximum daily amount of ≤10 g/day.

TABLE 1 Elemental Impurities for Drug Products Concentration Limits(mg/g) for Mucosal or Topical Product with a Element Maximum Daily Doseof ≤10 g/day Cadmium 2.5 Lead 0.5 Inorganic arsenic 0.15 Inorganicmercury 1.5 Iridium 10 Osmium 10 Palladium 10 Platinum 10 Rhodium 10Ruthenium 10 Molybdenum 10 Nickel 50 Vanadium 10 Copper 100 a PDE =Permissible daily exposure based on a 50-kg person.

Methods of detecting elemental impurities are known in the art andinclude inductively coupled plasma-atomic (optical) emissionspectroscopy (ICP-AES or ICP-OES) or ICP mass spectrometry.

The presence of certain microorganisms in non-sterile compositions mayhave the potential to reduce or even inactivate the therapeutic activityof a composition and may adversely affect the health of the patient.Good Manufacturing Practice may be used to ensure a low bioburden offinished dosage forms.

In certain embodiments, compositions disclosed herein adhere toacceptance criteria for nonsterile pharmaceutical products for topicaluse as set forth in Table 2, which are based upon the total aerobicmicrobial count (TAMC) and the total combined yeasts and molds count(TYMC).

TABLE 2 Acceptance Criteria for Microbiological Quality Total CombinedRoute of Total Aerobic Yeasts/Molds Adminis- Microbial Count CountSpecified tration (cfu/g or cfu/mL) (cfu/g or cfu/mL) Microorganism(s)Vaginal 100 10 Absence of Pseudomonas aeruginosa (1 g or 1 mL) Absenceof Staphylococcus aureus (1 g or 1 mL) Absence of Candida albicans (1 gor 1 mL)

Tests for determining whether a composition complies with aspecification for microbiological quality of a non-sterile compositionare known in the art (e.g., United States Pharmacopeia and The NationalFormulary (USP-NF), Chapter 61 and Chapter 62). In certain embodiments,compositions disclosed herein adhere to acceptance criteria for sterilepharmaceutical products. Sterilization of the composition may beachieved by methods known in the art, including heat (e.g., steam underpressure, filtration high pressure, irradiation (non-ionizing andionizing), and chemicals (e.g., ethylene oxide, nitrogen dioxide,formaldehyde). Tests for determining whether a composition complies witha specification for microbiological quality for a sterile compositionare known in the art (e.g., United States Pharmacopeia and The NationalFormulary (USP-NF), Chapter 71). In some embodiments, a sterilizationprocedure for a composition is validated to assure sterility of thecomposition and that no adverse change has occurred in the compositionas a result of the sterilization procedure. For example, a decrease inviscosity may occur in a composition following steam sterilization. Aformula for a composition may be modified to accommodatepost-sterilization changes in viscosity to retain or achieve desiredcomposition viscosity.

The compositions disclosed herein may be formulated as a semi-solid,gel, ointment, liquid, lotion, jelly, film, suspension, emulsion,mucilage, foam, cream, paste, or aerosol. In certain embodiments, thecompositions are formulated as a gel. The compositions may beadministered topically to the female lower reproductive tract (e.g., tothe vulva or into the vagina), to a sex toy that is to be inserted intothe vagina of a female subject, to the male penis prior to sexualintercourse or manual semen collection, or used to lubricate medicaldevices or instruments in the context of gynecological procedures orassisted reproduction techniques. The compositions of the presentdisclosure are formulated so as to allow the xylose, Salvia extract, andany other therapeutic agent contained therein to be bioavailable uponadministration of the composition to the subject.

The compositions of the present disclosure may be administered to asubject as a single dosage unit or the compositions may be administeredas a plurality of dosage units. For example, the compositions may besterilized, and packaged preservative-free in single-use, plasticlaminated pouches or plastic tubes of dimensions selected to provide forroutine, measured dispensing. In one example, a container may havedimensions used to dispense a single dose (e.g., about 4 ml) of thecomposition (e.g., a gel form) to the lower reproductive tract. Inanother example, compositions may comprise a paraben-free preservativeand packaged in a multi-use container used to dispense multiple doses ofthe composition.

In certain embodiments, compositions of this disclosure may beadministered daily (e.g., q.d., b.i.d., t.i.d., q.i.d), weekly, monthly,or on an as needed basis. In some embodiments, compositions of thepresent disclosure are administered in an amount of about 0.5 ml toabout 100 ml, about 1 ml to about 75 ml, about 2.5 ml to about 50 ml,about 5 ml to about 40 ml, about 10 ml to about 30 ml, about 15 ml toabout 25 ml, about 0.5 ml to about 50 ml, about 0.5 ml to about 40 ml,about 0.5 ml to about 30 ml, about 0.5 ml to about 20 ml, about 0.5 mlto about 10 ml, about 0.5 ml to about 5 ml, about 1 ml to about 50 ml,about 1 ml to about 40 ml, about 1 ml to about 30 ml, about 1 ml toabout 20 ml, about 1 ml to about 10 ml, or about 1 ml to about 5 ml. Insome embodiments, compositions of the present disclosure areadministered in an amount of about 0.25 ml, 0.5 ml, 0.75 ml, 1 ml, 2 ml,3 ml, 4 ml, 5 ml, 6 ml, 7 ml, 8 ml, 9 ml, 10 ml, 11 ml, 12 ml, 13 ml, 14ml, 15 ml, 16 ml, 17 ml, 18 ml, 19 ml, 20 ml, 25 ml, 30 ml, 35 ml, 40ml, 45 ml, 50 ml, 55 ml, 60 ml, 65 ml, 70 ml, 75 ml, 80 ml, 85 ml, 90ml, 95 ml, or 100 ml.

Any of the compositions disclosed herein may be used in methods forpromoting or enhancing vaginal ecosystem homeostasis of the lowerreproductive tract, comprising administering a composition according tothe present disclosure to the lower reproductive tract of a femalesubject. Moreover, any of the compositions disclosed herein may be usedin methods for increasing hydration of the lower reproductive tract,comprising administering a composition according to the presentdisclosure to the lower reproductive tract of a female subject.

Furthermore, the compositions disclosed herein may be used as a basecomposition in a variety of products directed to the female reproductivetract, which can be further modified according to the life stage (e.g.,reproductive age, menopause) or menstrual phase (e.g., ovulatory,luteal, pregnant) of the female subject and intended use of the product(e.g., perineal massage, contraceptive, anti-microbial, fertilityenhancement, sexual lubricant, vaginal hydration, sexual arousal,vaginal tightening, vaginal freshening, vaginal homeostasis, or drugcarrier base). Non-limiting examples of such products follow in furtherdetail.

In yet another aspect, the compositions disclosed herein may be usedduring sexual intercourse or artificial insemination by application tothe male penis prior to or during sexual intercourse or manualcollection of sperm, or to condoms (interior, exterior, or both) duringsexual intercourse, or to sex toys prior to use. Generally, sperm donorscollect semen samples by manual manipulation without the benefit oflubrication because available lubricants and saliva are spermicidal. Incertain embodiments, the compositions of the present disclosure arenon-toxic to sperm and may maintain, preserve, enhance, or increasesperm survival or function. In certain embodiments, the compositionsdisclosed herein may be applied directly to the penis, to the hand, coatthe interior or exterior of a condom, or be placed in a receptacle forsperm collection such as a vial, tube, baggie, or other collectiondevices.

In addition, the compositions disclosed herein may be used in variousassisted reproductive techniques, diagnostic procedures, and medicalprocedures. In certain embodiments, the compositions of the presentdisclosure are non-toxic to oocytes or embryos and may maintain,preserve, enhance, or increase oocyte or embryo survival or function.For example, a composition may be used to coat a speculum during pelvicexaminations, a catheter for insertion into a bladder to collect spermfrom a retrograde ejaculation, a uterine balloon tamponade, or a vaginalultrasound probe. A composition may be used to lubricate a catheter,pipette, hand, or other medical device or instrument in the context of avariety of assisted reproduction procedures including egg retrieval, eggfreezing, intracytoplasmic sperm injection (ICSI), embryo culture,embryo freezing, embryo thawing, embryo transfer, and artificialinsemination. A composition may also be used as a lubricant indiagnostic procedures such as endoscopy, hysteroscopy,sonohysterosalpingogram, contrast radiography (e.g.,hysterosalpingogram), or biopsy (e.g., endometrial biopsy). Thecompositions disclosed herein may be used in any variety of species forsperm collection, sexual intercourse, assisted reproductive techniques,diagnostic or medical procedures, and the like. Such subjects include,but are not limited to, humans, bovine, equine, canine, ovine, avian,feline, porcine, avian, rodent, lagomorph, caprine, primate, and variousexotic or rare species (e.g., elephant, lion, rhinoceros). Inembodiments where a LRT composition is used in the context of improvingfertility, assisted reproductive techniques, or in a female subject thatis in the ovulatory phase, the pH range of the composition is matched tothe LRT during the ovulatory phase (pH of about 6.8 to about 7.8). Inembodiments where a LRT composition is used in a medical or diagnosticcontext on a female subject who is in a non-ovulatory phase (forreproductive aged woman), in menopause, or pregnant, the pH range of thecomposition is matched to the LRT to the cycle or life specific phase(pH of about 3.5 to about 5.5, pH of about 4.5 to about 6.8, and pH ofabout 4.5 to about 6.5, respectively).

(1) Perineal Massage Compositions

In one aspect of the present disclosure, compositions formulated (e.g.,in the form of a gel) for use in perineal massage are provided. Thepractice of perineal massage (PM) widens and relaxes a woman's birthcanal during her last month of pregnancy. The perineum includes the backportion of a woman's birth canal (e.g. vagina and vulva). A perineumthat can't stretch to let the baby's head through the birth canal willundergo perineal trauma and either tear, or be cut by a doctor in anepisiotomy. These perineal tears or cuts require suturing to heal, whichcan often scar, causing pain after childbirth. This can negativelyimpact sexual function or lead to incontinence of urine or feces. Anintact perineum is one that stretches to let the baby out and doesn'tundergo tearing or cutting, and suturing. Studies have shown that womenwho keep an intact perineum during childbirth have less pain andincontinence and better sexual satisfaction post-partum. Nearly half ofwomen undergoing childbirth will suffer some perineal trauma. Perinealmassage in the final month of pregnancy decreases the rates of perinealtrauma, episiotomies and postpartum pain.

Perineal massage is a daily manual stretching of the back of the birthcanal, starting at 34 weeks of gestation for at least 4 or at least 5minutes each day. Current recommendations for perineal massage includeuse of aloe vera gel, almond oil, K-Y® Jelly or Vaseline for theprocedure. However, almond oil is known to have high levels of oxidativechemicals, which are potent triggers of the LRT inflammation system.K-Y® Jelly is known to be severely irritating and to cause the death ofvaginal microbiota. Aloe vera gels contain high levels of endotoxins andheavy metals, which can cause cell toxicity and inflammation. Vaselinehas been shown to cause an 8-fold increase in vaginal yeast infections.

The compositions disclosed herein that are pH matched to the vulvar andvaginal pH of pregnant women (about 4.5 to about 6.5) can be used as aperineal massage composition. In another embodiment, a perineal massagecomposition may also be used during labor to facilitate delivery of ababy during childbirth. The perineal massage compositions can beformulated as a semi-solid, gel, ointment, liquid, lotion, jelly, film,suspension, emulsion, mucilage, foam, cream, paste, or aerosol.

In certain embodiments, a perineal massage composition is used in amethod of reducing perineal trauma during child birth, comprisingadministering the perineal massage composition to the perineum, vagina,or external genitalia of a pregnant female subject. In some embodiments,the composition is gently massaged into the perineum, vagina, andexternal genitalia. In some embodiments, the perineal massage gel isadministered to the subject beginning at 34-36 weeks of gestation, andmay continue daily, every other day, 2-3 times a week, or weekly. Theperineal massage may continue for at least 1 minute, at least 2 minutes,at least 3 minutes, at least 4 minutes, or at least 5 minutes at a time.In some embodiments, the perineal massage composition may beadministered to the perineum, vagina, or external genitalia of thesubject during labor.

(2) Microbicide or Contraceptive Compositions

In another aspect of the present disclosure, compositions that optimizethe formation of thick, hostile cervical mucus to decrease sperm and/orpathogen penetration through the vaginal and cervical canal areprovided. Over the counter, vaginally administered contraceptives andmicrobicides contain numerous inactive ingredients that may causeprofound mucosal irritation, epithelial toxicity, vaginal microbiotadamage, and interleukin secretion. Contraceptive active components andmicrobicides are de facto cell toxic compounds. Prior development ofvaginal drugs has unfortunately coupled such cytotoxic active drugs withhighly cytotoxic inactive carrier ingredients. Available commercialvaginal contraceptives include at least three inactive ingredients knownto cause significant LRT damage on their own, in addition to thecytotoxic contraceptive active ingredient, nonoxyonol-9. Existingnon-oxynol products have been shown to increase STD transmission. Thecompositions of the present disclosure provide LRT carrier formulas thatmaximize safety of such products.

The microbicide or contraceptive containing compositions provided hereinare matched to the vulvar and vaginal pH of the subject, e.g., for areproductive aged woman (about 3.5 to about 5.5), an ovulatory phasewoman (about 6.8 to about 7.8), or postmenopausal woman (around about4.5 to about 6.8).

In certain embodiments, the contraceptive compositions may impede spermfunction and transport through cervical mucus. In certain embodiments,the microbicide compositions stimulate increased clearance of pathogensand/or lower infection rates after exposure. In some embodiments, thecompositions further comprise an additional contraceptive agent oranti-microbial agent (e.g., antibiotic, anti-fungal agent, anti-viralagent, or any combination thereof). A microbicide or contraceptivecomposition can be formulated as a semi-solid, gel, ointment, liquid,lotion, jelly, film, suspension, emulsion, mucilage, foam, cream, paste,or aerosol. A microbicide or contraceptive composition can be used as alubricant, contraceptive, anti-microbial, drug carrier base, for vaginalhydration, vaginal freshness, vaginal homeostasis, or any combinationthereof.

An exemplary formulation of a contraceptive composition X and % rangesof each component as contemplated in the present disclosure is set forthas follows: 85.248% by weight purified water, 1.0% by weight cetylhydroxyethylcellulose, 2.0% by weight carbomer, 0.139% by weight lacticacid, 0.683% by weight sodium lactate, 0.232% by weight phenethylalcohol, 0.168% by weight caprylyl glycol, 0.05% by weight Salviasclarea, 0.15% by weight sodium chloride, 0.05% by weight calciumchloride, 10% by weight Salvia hispanica, 0.04% by weight d-Xylose, and0.1%-1% sodium hydroxide. In certain embodiments, contraceptivecomposition X is formulated as a gel.

In certain embodiments, lower doses of contraceptive drugs oranti-microbial drugs may be used in the compositions if the base formulaof the composition thickens cervical, mucus, impairs sperm function, orimpairs microbial penetration of the cervical mucus.

In certain embodiments, the microbicide containing compositions may beused in a method of treating an infection of the lower reproductivetract, comprising administering the microbicide containing compositionto the lower reproductive tract of a female subject (e.g.,intravaginally, to the vulva).

In certain embodiments, the contraceptive containing compositions may beused in a method of inhibiting conception, comprising administering thecontraceptive containing composition intravaginally, to the vulva orpenis, to condoms (interior or exterior), or other devices orinstruments. In some embodiments, the contraceptive containingcomposition is administered within 24 hours, 12 hours, 6 hours, 2 hours,1 hour, 30 minutes, 15 minutes, 5 minutes, just prior to, or duringsexual intercourse.

(3) Fertility Compositions

In another aspect of the present disclosure, fertility compositions areprovided. During the ovulatory phase of the menstrual cycle, vaginallyapplied compositions may be used to support and enhance penetration ofthe cervical mucus by sperm. Current commercial vaginal fertilityproducts contain ingredients that trigger interleukin production andepithelial disruption of the vaginal mucosa. Commercial vaginalfertility products also contain ingredients that are physiologicallydecreased during ovulation in vivo (e.g., fructose and calcium),ingredients that may compromise cervical mucus and vaginal secretionhydration, possibly impairing sperm transport to the egg. At present,there are currently no vaginal products available to enhance quality offertile cervical mucus, to supplement glycosaminoglycans and mucinproduction levels during ovulation, and to support hydration of thecervical mucus granules and fertile vaginal secretions, as needed tooptimize sperm transport. Further, there are no vaginal fertilityproducts that are free of mucosal immune-stimulating excipients.

Fertility compositions of the present disclosure (e.g., FertilityComposition Formulas X, Y, Z, AZ, AB, AB′, and AC) can be used for:fertility enhancement, lubrication, vaginal hydration, sexual arousal,vaginal tightening, vaginal freshening, vaginal homeostasis, drugcarrier base, or any combination thereof.

In another aspect, the fertility compositions of the present disclosuremay serve to hydrate the vaginal mucosa of ovulating women withoutharming VMB, as an alternative means to optimizing sperm transport, oras an additional method for optimizing natural fertility.

Fertility compositions of this disclosure for use in ovulating women arepH matched to the vaginal pH of the ovulatory phase female subject,about pH 6.8 to about 7.8. In certain embodiments, a fertilitycomposition has a pH of about 7.0 to about 7.6, of about 7.0 to about7.4, of about 7.0 to about 7.2, or about 7.0.

In further embodiments, a fertility composition is formulated to benon-toxic to sperm (non-spermicidal) and may maintain, preserve,enhance, improve, or increase sperm survival and function, e.g.,penetration into the fertile cervical mucus of an ovulating woman orimproved potential of sperm to fertilize an oocyte. In yet furtherembodiments, a fertility composition may improve cervical mucus andfertile vaginal fluid hydration and quality to optimize sperm transport.

In yet further embodiments, a fertility composition is formulated to benon-toxic to oocytes or embryos and may maintain, preserve, enhance,improve, or increase oocyte or embryo survival and function, e.g.,oocyte maturation, oocyte potential for fertilization by sperm, embryopotential for blastocyst development.

In certain embodiments, fertility compositions of this disclosure areused in a method of enhancing fertility, comprising administering afertility composition intravaginally, to the vulva, or penis. In someembodiments, the fertility composition is administered within 24 hours,12 hours, 6 hours, 2 hours, 1 hour, 30 minutes, 15 minutes, 5 minutes,just prior to, or during sexual intercourse.

In another aspect, the fertility compositions of the present disclosureare used in an assisted reproduction setting. For example, it may beapplied to the hand, condom (interior or exterior), penis, or othercollection device (e.g., vial, tube, bag) for use in semen collectionfor analysis, freezing, or use in artificial insemination (e.g.,intrauterine insemination) or in vitro fertilization techniques (e.g.,ICSI). In another example, the fertility compositions of the presentdisclosure may be applied to a medical device or instrument (e.g.,catheter, pipette), or storage container for use in sperm collection(manual or surgical), artificial insemination, oocyte retrieval, oocytefreezing, oocyte thawing, ICSI, embryo culture, embryo biopsy, embryofreezing, embryo thawing, or embryo transfer.

In other embodiments, fertility compositions of the present disclosureare used in a method of increasing hydration of the vaginal mucosacomprising administering a fertility composition to the lowerreproductive tract of a female subject. In some embodiments, the femalesubject is in the ovulatory phase of the menstrual cycle.

Fertility compositions of the present disclosure can be formulated as asemi-solid, gel, ointment, liquid, lotion, jelly, film, suspension,emulsion, mucilage, foam, cream, paste, or aerosol.

Exemplary formulations of fertility compositions and % ranges of eachcomponent as contemplated in the present disclosure are set forth asfollows.

Fertility Composition Formula W comprises: 96.5% by weight purifiedwater (range 50%-99%); 2% by weight carboxymethylcellulose (range0.1%-10%); 0.01% by weight vascular endothelial growth factor (range0.001%-1%); 0.2% by weight monobasic sodium phosphate (range 0.01%-2%);0.7% by weight dibasic potassium phosphate (range 0.01%-2%); 0.25% byweight carvacrol (range 0.01%-0.5%); 0.1% by weight Salvia plebeia(range 0.001%-5%); 0.4% by weight sodium chloride (range 0.01%-1%);0.04% xylose (range 0.01%-5%); and 0.001%-1% by weight sodium hydroxide.In certain embodiments, Fertility Composition Formula W is formulated asa gel.

Fertility Composition Formula X comprises: 94.81% by weight purifiedwater (range 50%-99%); 1.4% by weight polyvinyl alcohol (range0.01%-5%); 1.5% by weight hypromellose (range 0.1%-10%); 1.46% by weightdibasic sodium phosphate (range 0.01%-2%); 0.47% by weight potassiumcitrate (range 0.01%-2%); 0.01% by weight oleuropein (range 0.01%-2%);0.05% by weight Salvia sclarea (range 0.01%-1%); 0.2% by weight sodiumchloride (range 0.01%-1%); 0.1% by weight xylose (range 0.01%-5%); and0.001%-1% by weight sodium hydroxide. In certain embodiments, FertilityComposition Formula X is formulated as a gel.

Fertility Composition Formula Y comprises: 88.75% by weight purifiedwater (range 50%-99%); 0.4% by weight poloxamer (range 0.1%-5%); 0.2% byweight monobasic sodium phosphate (range 0.01%-2%); 0.7% by weightdibasic potassium phosphate (range 0.01%-2%); 0.05% by weight Salviasclarea (range 0.01%-1%); 0.4% by weight sodium chloride (range0.01%-1%); 0.25% by weight carvacrol (range 0.01%-0.5%); 9% by weightPoly(2-methyl-2-oxazoline) (range 0.5%-40%); and 0.001%-1% by weightsodium hydroxide. In certain embodiments, Fertility Composition FormulaY is formulated as a gel.

Fertility Composition Formula Z comprises: 96.81% by weight purifiedwater (range 50%-99%); 0.7% by weight hypromellose (range 0.1%-10%);0.27% by weight carbomer (range 0.1%-5%); 1.46% by weight dibasic sodiumphosphate (range 0.01%-2%); 0.47% by weight potassium citrate (range0.01%-2%); 0.3% by weight phenoxyethanol (0.1%-2%); 0.2% by weightpotassium sorbate (range 0.025%-1%); 0.05% by weight Salvia sclarea(range 0.01%-1%); 0.4% by weight sodium chloride (range 0.01%-1%); 0.04%by weight xylose (range 0.01%-5%); 0.001%-1% by weight sodium hydroxide.In certain embodiments, Fertility Composition Formula Z is formulated asa gel.

Fertility Composition Formula AZ comprises: 88.69% by weight purifiedwater (range 50%-99%); 0.1% by weight Salvia plebeia (range 0.01%-1%);0.1% by weight phenoxyethanol (range 0.1%-2%); 0.3% by weight caprylylglycol (range 0.01%-2%); 0.5% by weight sodium chloride (range0.01%-1%); 0.2% by weight monobasic potassium phosphate (range0.01%-2%); 0.7% by weight dibasic sodium phosphate (range 0.01%-2%);0.4% by weight xylose (range 0.01%-5%); 9% by weightpoly(2-methyl-2-oxazoline) (range 0.5%-40%); and 0.001%-1% by weightsodium hydroxide. In certain embodiments, Fertility Composition FormulaAZ is formulated as a gel.

Fertility Composition Formula AB comprises: 96.98% by weight purifiedwater (range 75%-99.5%); 0.7% by weight cetyl hydroxyethylcellulose(range 0.001%-25%); 0.5% by weight hypromellose (range 0.001%-25%);0.25% by weight carbomer (range 0.001%-20%); 0.188% by weight monobasicsodium phosphate (range 0.02%-2%); 0.687% by weight dibasic potassiumphosphate (range 0.07%-7%); optionally 0.232% by weight phenethylalcohol (0.02%-2%); optionally 0.148% by weight caprylyl glycol (range0.01%-2%); 0.05% by weight Salvia sclarea (range 0.005%-1%); 0.2% byweight sodium chloride (range 0.001%-5%); 0.04% by weight xylose (range0.001%-5%); and 0.25% by weight sodium hydroxide (range 0.001%-1%). Incertain embodiments, Fertility Composition Formula AB is formulated as agel.

Fertility Composition Formula AB′ comprises: 96.24% by weight purifiedwater (range 85%-98%); 0.7% by weight cetyl hydroxyethylcellulose (range0.1%-2%); 0.9% by weight hypromellose (range 0.1%-2%); 0.27% by weightcarbomer (range 0.1%-1%); 0.188% by weight monobasic sodium phosphate(range 0.1%-1%); 0.687% by weight dibasic potassium phosphate (range0.1%-1%); optionally 0.232% by weight phenethyl alcohol (0.05%-1%);optionally 0.168% by weight caprylyl glycol (range 0.05%-1%); 0.05% byweight Salvia sclarea (range 0.01%-2.5%); 0.4% by weight sodium chloride(range 0.1%-1%); 0.04% by weight xylose (range 0.0015%-2.5%); and 0.12%by weight sodium hydroxide (range 0.001%-1%). In certain embodiments,Fertility Composition Formula AB′ is formulated as a gel.

Fertility Composition Formula AC comprises: 97% by weight purified water(range 95%-99%); 1% by weight hypromellose (range 0.1%-2%); 0.5% byweight hydroxyethyl cellulose (range 0.1%-1%); 0.2% by weight monobasicsodium phosphate (range 0.02%-2%); 0.7% by weight dibasic potassiumphosphate (range 0.07%-7%); 100,000 cfu Lactobacillus sp. (range 10,000cfu-1,000,000 cfu); 0.464% by weight phenethyl alcohol (range 0.02%-2%);0.336% by weight caprylyl glycol (range 0.01%-2%); 0.05% by weightSalvia sclarea (range 0.005%-1%); 0.12% by weight sodium chloride (range0.001%-5%); 0.04% by weight xylose (range 0.001%-5%); and 0.001% byweight sodium hydroxide (range 0.001%-1%). In certain embodiments,Fertility Composition Formula AC is formulated as a gel.

In certain embodiments, topical, isotonic, biome-friendly fertilitycompositions of the present disclosure comprise xylose at aconcentration ranging from about 0.01% to about 2.5% by weight and aSalvia extract at a concentration ranging from about 0.01% to about 2.5%by weight, wherein the composition has a pH of about 6.8 to about 7.8and is optionally formulated as a gel. In some embodiments, thefertility composition comprises xylose at a concentration ranging fromabout 0.02% to about 0.5% by weight and Salvia extract at aconcentration ranging from about 0.01% to about 0.2% by weight. In someembodiments, the Salvia extract is Salvia sclarea, Salvia hispanica,Salvia plebeia, or combinations thereof.

The fertility compositions may further comprise a non-irritatingviscosity increasing agent at a concentration ranging from about 0.1% toabout 25% by weight. Examples of non-irritating viscosity-increasingagent include a cellulose ether (e.g., methyl cellulose, carboxymethylcellulose, hypromellose, ethylcellulose, ethyl methyl cellulose,hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxyethyl methylcellulose, ethyl hydroxyethyl cellulose, or a combination thereof)carbomer, polyoxazoline, or any combination thereof. In someembodiments, non-irritating viscosity-increasing agent is hypromelloseand carbomer homopolymer type B together at a total concentrationranging from about 0.5% to about 5% by weight. In further embodiments,the ratio of hypromellose to carbomer homopolymer type B in thecomposition ranges from about 2:1 to about 6:1. In some embodiments, thenon-irritating viscosity-increasing agent is hypromellose, a carbomer,carobxymethylcellulose, or poly(2-methyl-2-oxazoline).

The fertility compositions may further comprise a surfactant at a totalconcentration ranging from about 0.1% to about 2% by weight. Examples ofsurfactants include cetyl hydroxyethylcellulose, polyvinyl alcohol,poloxamer, and any combination thereof. In some embodiments, thesurfactant is a cetyl hydroxyethylcellulose at a total concentrationranging from about 0.35% to about 2% by weight. In other embodiments,the surfactant is a polyvinyl alcohol at a total concentration rangingfrom about 1% to about 1.5% by weight. In yet other embodiments, thesurfactant is a poloxamer at a total concentration ranging from about0.1% to about 0.5% by weight.

The fertility compositions may include a paraben-free preservative at atotal concentration ranging from about 0.005% to 5% by weight. In someembodiments, the paraben-free preservative is a phenethyl alcohol,caprylyl glycol, phenoxyethanol, a sorbate, oleuropein, acemannan,carvacrol, or any combination thereof. In yet further embodiments, theparaben-free preservative is a phenethyl alcohol and caprylyl glycoltogether at a total concentration ranging from about 0.25% to about 1%by weight. In other embodiments, the paraben-free preservative is aphenoxyethanol and caprylyl glycol at a total concentration ranging fromabout 0.25% to about 1% by weight. In yet other embodiments, theparaben-free preservative is a phenoxyethanol and a sorbate, wherein thesorbate is a potassium sorbate, at a total concentration ranging fromabout 0.4% to about 1% by weight. In yet other embodiments, theparaben-free preservative is oleuropein. In yet other embodiments, theparaben-free preservative is a carvacrol at a total concentrationranging from about 0.01% to about 0.5% by weight.

The fertility compositions may further comprise a buffering agent at atotal concentration ranging from about 0.5% to about 2.5% by weight. Incertain embodiments, the buffering agent is sodium phosphate (monobasic,dibasic, or both), potassium phosphate (monobasic, dibasic, or both),potassium citrate, or any combination thereof. In some embodiments, thebuffering agent is sodium phosphate (monobasic, dibasic, or both) andpotassium phosphate (monobasic, dibasic, or both) at a totalconcentration ranging from about 0.5% to about 1.5% by weight. In otherembodiments, the buffering agent is sodium phosphate and potassiumcitrate at a total concentration ranging from about 1% to about 2% byweight.

The fertility compositions may also comprise a pH modifying agent at atotal concentration ranging from about 0.001% to about 2% by weight. ThepH modifying agent can be an acidifying agent, an alkalizing agent, orboth. In certain embodiments, the alkalizing agent is ammonia, ammoniumcarbonate, diethanolamine, monoethanolamine, potassium hydroxide,dibasic potassium phosphate, sodium bicarbonate, sodium borate, sodiumcarbonate, sodium hydroxide, dibasic sodium phosphate, trolamine, or anycombination thereof. In some embodiments, the alkalizing agent is sodiumhydroxide at a total concentration ranging from about 0.01% to about 1%by weight.

The fertility compositions may further comprise an osmolality adjuster,such as sodium chloride, potassium chloride, or both, at a totalconcentration ranging from about 0.1% to about 1% by weight.

The fertility compositions may further comprise a solvent, such as waterin an amount of about 85% to about 98% by weight.

An exemplary topical, isotonic, biome-friendly fertility compositioncomprises xylose at a concentration ranging from about 0.01% to about2.5% by weight or about 0.02% to about 0.5% by weight and Salvia plebeiaat a concentration ranging from about 0.05% to about 0.15% by weight,and the composition further comprises: a non-irritatingviscosity-increasing agent, wherein the non-irritatingviscosity-increasing agent is carboxymethyl cellulose at a totalconcentration ranging from about 1% to about 3% by weight; a vascularendothelial growth factor at a total concentration ranging from about0.005% to about 0.05% by weight; a buffering agent, wherein thebuffering agent is monobasic sodium phosphate and dibasic potassiumphosphate together at a total concentration ranging from about 0.5% toabout 1.1% by weight; an osmolality adjuster, wherein the osmolalityadjuster is sodium chloride at a total concentration ranging from about0.1% to about 0.7% by weight; a solvent, wherein the solvent is water ata concentration ranging from about 95% to about 98% by weight; and a pHmodifier, wherein the pH modifier is sodium hydroxide at a concentrationranging from about 0.001% to about 1% by weight, and optionally, aparaben-free preservative, wherein the paraben-free preservative ispreferably carvacrol at a total concentration ranging from about 0.01%to about 0.5% by weight, wherein the composition has a pH of about 6.8to about 7.8. The fertility composition can be formulated as a gel.

Another exemplary topical, isotonic, biome-friendly fertilitycomposition comprises xylose at a concentration ranging from about 0.01%to about 2.5% by weight or about 0.02% to about 0.5% by weight andSalvia plebeia at a concentration ranging from about 0.05% to about0.15% by weight, and the composition further comprises: a non-irritatingviscosity-increasing agent, wherein the non-irritatingviscosity-increasing agent is poly(2-methyl-2-oxazoline) at a totalconcentration ranging from about 5% to about 10%; a buffering agent,wherein the buffering agent is monobasic potassium phosphate and dibasicsodium phosphate together at a total concentration ranging from about0.5% to about 1.1% by weight; an osmolality adjuster, wherein theosmolality adjuster is sodium chloride at a total concentration rangingfrom about 0.1% to about 0.75% by weight; a solvent, wherein the solventis water at a concentration ranging from about 87% to about 94% byweight; a pH modifier, wherein the pH modifier is sodium hydroxide at aconcentration ranging from about 0.001% to about 1% by weight; andoptionally, a paraben-free preservative, wherein the paraben-freepreservative is preferably a phenoxyethanol and caprylyl glycol togetherat a total concentration ranging from about 0.1% to about 0.7% byweight, and wherein the composition has a pH of about 6.8 to about 7.8.The fertility composition can be formulated as a gel.

Another example of a topical, isotonic, biome-friendly fertilitycomposition comprises xylose at a concentration ranging from about 0.01%to about 2.5% by weight or about 0.02% to about 0.5% by weight andSalvia sclarea at a concentration ranging from about 0.025% to about0.075% by weight, and the composition further comprises: anon-irritating viscosity-increasing agent, wherein the non-irritatingviscosity-increasing agent is hypromellose at a total concentrationranging from about 1% to about 2% by weight; a surfactant, wherein thesurfactant is polyvinyl alcohol at a total concentration ranging fromabout 1% to about 2% by weight; a buffering agent, wherein the bufferingagent is dibasic sodium phosphate and potassium citrate together at atotal concentration ranging from about 1% to about 3% by weight; anosmolality adjuster, wherein the osmolality adjuster is sodium chlorideat a total concentration ranging from about 0.1% to about 0.3% byweight; a solvent, wherein the solvent is water at a concentrationranging from about 95% to about 98% by weight; a pH modifier, whereinthe pH modifier is sodium hydroxide at a concentration ranging fromabout 0.001% to about 1% by weight; and optionally, a paraben-freepreservative, wherein the paraben-free preservative is preferablyoleuropein at a total concentration ranging from about 0.01% to about0.1% by weight, and wherein the composition has a pH of about 6.8 toabout 7.8. The fertility composition can be formulated as a gel.

Yet another exemplary topical, isotonic, biome-friendly fertilitycomposition comprises xylose at a concentration ranging from about 0.01%to about 2.5% by weight or about 0.02% to about 0.5% by weight andSalvia sclarea at a concentration ranging from about 0.025% to about0.075% by weight, and the composition further comprises: anon-irritating viscosity-increasing agent, wherein the non-irritatingviscosity-increasing agent is poly(2-methyl-2-oxazoline) at a totalconcentration ranging from about 5% to about 10% by weight; asurfactant, wherein the surfactant is a poloxamer at a totalconcentration ranging from about 0.1% to about 0.7% by weight; abuffering agent, wherein the buffering agent is monobasic sodiumphosphate and dibasic potassium phosphate together at a totalconcentration ranging from about 0.5% to about 1.1% by weight; anosmolality adjuster, wherein the osmolality adjuster is sodium chlorideat a total concentration ranging from about 0.1% to about 0.5% byweight; a solvent, wherein the solvent is water at a concentrationranging from about 87% to about 94% by weight; a pH modifier, whereinthe pH modifier is sodium hydroxide at a concentration ranging fromabout 0.001% to about 1% by weight; and optionally, a paraben-freepreservative, wherein the paraben-free preservative is preferablycarvacrol at a total concentration ranging from about 0.03% to about0.5% by weight, and wherein the composition has a pH of about 6.8 toabout 7.8. The fertility composition can be formulated as a gel.

Another example of a topical, isotonic, biome-friendly fertilitycomposition comprises xylose at a concentration ranging from about 0.01%to about 2.5% by weight or about 0.02% to about 0.5% by weight andSalvia sclarea at a concentration ranging from about 0.025% to about0.075% by weight, and the composition further comprises: anon-irritating viscosity-increasing agent, wherein the non-irritatingviscosity-increasing agent is hypromellose and carbomer homopolymer typeB together at a total concentration ranging from about 0.2% to about 3%by weight, and wherein the ratio of hypromellose to carbomer homopolymertype B in the composition is about 2:1; a surfactant, wherein thesurfactant is cetyl hydroxyethylcellulose at a total concentrationranging from about 0.5% to about 1% by weight; a buffering agent,wherein the buffering agent is monobasic sodium phosphate and dibasicpotassium phosphate together at a total concentration ranging from about0.5% to about 1.5% by weight; an osmolality adjuster, wherein theosmolality adjuster is sodium chloride at a total concentration rangingfrom about 0.1% to about 0.7% by weight; a solvent, wherein the solventis water at a concentration ranging from about 95% to about 98% byweight; a pH modifier, wherein the pH modifier is sodium hydroxide at aconcentration ranging from about 0.001% to about 1% by weight; andoptionally, a paraben-free preservative, wherein the paraben-freepreservative is preferably a phenethyl alcohol and caprylyl glycoltogether at a total concentration ranging from about 0.2% to about 0.9%by weight, and wherein the composition has a pH of about 6.8 to about7.8. The fertility composition can be formulated as a gel.

Another example of a topical, isotonic, biome-friendly fertilitycomposition comprises xylose at a concentration ranging from about 0.01%to about 2.5% by weight or about 0.02% to about 0.5% by weight andSalvia sclarea at a concentration ranging from about 0.025% to about0.075% by weight, and the composition further comprises: anon-irritating viscosity-increasing agent, wherein the non-irritatingviscosity-increasing agent is hypromellose and carbomer homopolymer typeB together at a total concentration ranging from about 0.2% to about 2%by weight, and wherein the ratio of hypromellose to carbomer homopolymertype B in the composition is about 3.3:1; a surfactant, wherein thesurfactant is cetyl hydroxyethylcellulose at a total concentrationranging from about 0.5% to about 1% by weight; a buffering agent,wherein the buffering agent is monobasic sodium phosphate and dibasicpotassium phosphate together at a total concentration ranging from about0.5% to about 1.5% by weight; an osmolality adjuster, wherein theosmolality adjuster is sodium chloride at a total concentration rangingfrom about 0.1% to about 0.7% by weight; a solvent, wherein the solventis water at a concentration ranging from about 95% to about 98% byweight; a pH modifier, wherein the pH modifier is sodium hydroxide at aconcentration ranging from about 0.001% to about 1% by weight; andoptionally, a paraben-free preservative, wherein the paraben-freepreservative is preferably a phenethyl alcohol and caprylyl glycoltogether at a total concentration ranging from about 0.2% to about 0.9%by weight, and wherein the composition has a pH of about 6.8 to about7.8. The fertility composition can be formulated as a gel.

Yet another exemplary topical, isotonic, biome-friendly fertilitycomposition comprises xylose at a concentration ranging from about 0.01%to about 2.5% or about 0.02% to about 0.5% by weight and Salvia sclareaat a concentration ranging from about 0.025% to about 0.075% by weight,and the composition further comprises: a non-irritatingviscosity-increasing agent, wherein the non-irritatingviscosity-increasing agent is hypromellose and carbomer homopolymer typeB together at a total concentration ranging from about 0.75% to about1.5% by weight, and wherein the ratio of hypromellose to carbomerhomopolymer type B in the composition is about 2.6:1; a buffering agent,wherein the buffering agent is dibasic sodium phosphate and potassiumcitrate together at a total concentration ranging from about 1% to about3% by weight; an osmolality adjuster, wherein the osmolality adjuster issodium chloride at a total concentration ranging from about 0.1% toabout 0.7% by weight; a solvent, wherein the solvent is water at aconcentration ranging from about 95% to about 98% by weight; a pHmodifier, wherein the pH modifier is sodium hydroxide at a concentrationranging from about 0.001% to about 1% by weight; and optionally, aparaben-free preservative, wherein the paraben-free preservative ispreferably a phenoxyethanol and potassium sorbate together at a totalconcentration ranging from about 0.25% to about 0.75% by weight, andwherein the composition has a pH of about 6.8 to about 7.8. Thefertility composition can be formulated as a gel.

Another exemplary topical, isotonic, biome-friendly fertilitycomposition comprises xylose at a concentration ranging from about 0.01%to about 2.5% or about 0.02% to about 0.5% by weight and Salvia sclareaat a concentration ranging from about 0.025% to about 0.075% by weight,and the composition further comprises: a non-irritatingviscosity-increasing agent, wherein the non-irritatingviscosity-increasing agent is hydroxyethyl cellulose and hypromellose ata total concentration ranging from about 0.2% to about 3%; a bufferingagent, wherein the buffering agent is monobasic sodium phosphate anddibasic potassium phosphate together at a total concentration rangingfrom about 0.5% to about 1.5% by weight; a Lactobacillus species that iscapable of colonizing the vaginal wall in an amount of about 10,000 cfuto about 1,000,000 cfu; an osmolality adjuster, wherein the osmolalityadjuster is sodium chloride at a total concentration ranging from about0.1% to about 0.7% by weight; a solvent, wherein the solvent is water ata concentration ranging from about 95% to about 99% by weight; a pHmodifier, wherein the pH modifier is sodium hydroxide at a concentrationranging from about 0.001% to about 1% by weight; and optionally, aparaben-free preservative, wherein the paraben-free preservative ispreferably a phenethyl alcohol and caprylyl glycol together at a totalconcentration ranging from about 0.2% to about 0.9% by weight, andwherein the composition has a pH of about 6.8 to about 7.8. Thefertility composition can be formulated as a gel.

(4) Sexual Lubricant and Vaginal Hydration Compositions

In yet another aspect of the present disclosure, compositions areprovided that may be used as sexual lubricants or for hydrating thevagina, or both. Lubricants alleviate discomfort or pain duringintercourse when vaginal mucosal secretions are inadequate. Inadequatevaginal secretion may occur due to lack of arousal, aging (menopause),as a side effect to medications (such as antidepressants and birthcontrol pills), or hormone disruptions. Over 70% of all American womenhave used a sexual lubricant at some time in their lives. However, thereare few lubricants on the market that lack ingredients shown to causeirritation or cytokine release from LRT cells or high levels ofinflammatory endotoxins. The most recent lubricants touted as“break-through” for women with vaginal dryness have been the hyaluronicacid (HA) based lubricants. However, the presence of elevated HA is aphysiologic trigger for inflammasome production in the human body.Additionally, HA production systems result in high levels of endotoxinand bacterial nucleic acids.

The sexual lubricant or vaginal hydration compositions of the presentdisclosure are matched to the vulvar and vaginal pH of the subject,e.g., for a reproductive aged woman (about pH 3.5 to about 5.5), anovulatory phase woman (about pH 6.8 to about 7.8), pregnant woman (aboutpH 4.5 to about 6.5), or postmenopausal woman (about. pH 4.5 to about6.8).

The compositions for lubrication or hydration of vaginal fluids and/ormucosa provided herein (e.g., Menopause Lubricant Composition FormulasW, X, Y, Z, AB) can be used for perineal massage, contraception,anti-microbial, fertility enhancement, sexual lubrication, vaginalhydration, sexual arousal, vaginal tightening, vaginal freshening,vaginal homeostasis, as a drug carrier base, or any combination thereof.

In certain embodiments, the hydration/lubricant compositions may includea therapeutic agent, such as a hormone useful for treating genitourinarysyndrome of menopause (e.g., estradiol, estriol, estropipate,testosterone, progesterone, DHEA, or a combination thereof) or othernatural products or botanicals that support hormone production. Incertain embodiments, the sexual hydration/lubricant compositions furthercomprise an agent that enhances vasodilation. In some embodiments, anagent that enhances vasodilation is L-arginine, niacin, nicotinamide,alprostadil, a phosphodiesterase inhibitor, or a combination thereof.

In certain embodiments, sexual lubricant compositions may be used inmethods of enhancing arousal during sexual intercourse, comprisingtopically applying a sexual lubricant composition to the lowerreproductive tract (e.g., intravaginally, to the vulva) of a femalesubject, to the penis, to condoms (interior or exterior), or to otherdevices (e.g., sex toys). In some embodiments, sexual lubricantcompositions are administered within 24 hours, 12 hours, 6 hours, 2hours, 1 hour, 30 minutes, 15 minutes, 5 minutes, just prior to, orduring sexual intercourse.

In certain embodiments, vaginal hydration compositions may be used inmethods of increasing hydration of the vaginal mucosa or secretions,comprising topically applying a vaginal hydration composition to thelower reproductive tract (e.g., intravaginally, to the vulva) of afemale subject or to the penis, condoms, or other devices or instruments(e.g., catheter, speculum, vaginal ultrasound probe, uterine balloontamponade) prior to insertion to the vagina.

Lubricant/vaginal hydration compositions of the present disclosure canbe formulated as a semi-solid, gel, ointment, liquid, lotion, jelly,film, suspension, emulsion, mucilage, foam, cream, paste, or aerosol.

Exemplary formulations of compositions for vaginal hydration orlubrication in menopausal subjects (“menopause compositions”) and %ranges of each component as contemplated in the present disclosure areset forth as follows. Such menopause compositions are pH matched to thevulvar and vaginal pH of post-menopausal subjects (about pH 4.5 to about6.8).

Menopause Composition Formula W comprises: 96.34% by weight purifiedwater (range 50-99%); 0.04% by weight xylose (range 0.01%-5%); 0.91% byweight poloxamer (range 0.1%-5%); 0.12% by weight benzoic acid (range0.003%-5%); 0.14% by weight sodium chloride (range 0.01%-1%); 1.0% byweight hypromellose (range 0.1%-10%); 0.05% by weight Salvia sclarea(range 0.1%-1%); 0.7% by weight monobasic sodium phosphate (range0.07%-7%); 0.135% by weight dibasic sodium phosphate (range 0.01%-1.4%);1.4% by weight polyvinyl alcohol (range 0.01%-5%); and 0.001%-1% byweight sodium hydroxide. In certain embodiments, Menopause CompositionFormula W is formulated as a gel.

Menopause Composition Formula X comprises: 96.14% by weight purifiedwater (range 50%-99%); 0.04% by weight xylose (range 0.01%-5%); 0.9% byweight poloxamer (range 0.1%-5%); 0.7% by weight monobasic sodiumphosphate (range 0.07%-7%); 0.135% dibasic sodium phosphate (range0.01%-1.4%); 0.03% by weight cranberry extract (range 0.01%-0.5%); 0.14%by weight sodium chloride (range 0.01%-1%); 2.0% by weight hypromellose(range 0.1%-10%); and 0.05% by weight Salvia sclarea (range 0.01%-1%).In certain embodiments, Menopause Composition Formula X is formulated asa gel.

Menopause Composition Formula Y comprises: 98.0% by weight purifiedwater (range 50%-99%); 0.04% by weight xylose (range 0.01%-5%); 0.7% byweight monobasic sodium phosphate (0.07%-7%); 0.135% by weight dibasicsodium phosphate (0.01%-1.4%); 0.232% by weight phenethyl alcohol (range0.01%-1%); 0.148% by weight caprylyl glycol (range 0.01%-2%); 0.14% byweight sodium chloride (range 0.01%-1%); 0.14% by weight carbomer (range0.01%-5%); 0.42% by weight hypromellose (range 0.1%-10%); 0.05% byweight Salvia plebeia (range 0.01%-1%); and 0.005% by weight sodiumhydroxide. (range 0.001%-1%) In certain embodiments, MenopauseComposition Formula Y is formulated as a gel.

Menopause Composition Formula Z comprises: 95.7% by weight purifiedwater (range 50%-99%); 0.1% by weight xylose (range 0.01%-5%); 1.07% byweight sodium citrate (range 0.01%-2%); 0.2% by weight potassium sorbate(range 0.025%-1%); 0.15% by weight sodium benzoate (range 0.1%-10%);0.24% by weight sodium chloride (range 0.01%-1%); 1.3% by weighthypromellose (range 0.1%-10%); 1.0% by weight carboxymethylcellulose(range 0.1%-10%); 0.05% by weight Salvia sclarea (range 0.01%-1%); 0.17%by weight citric acid (range 0.02%-20%); and 0.001% by weight sodiumhydroxide (range 0.001%4%). In certain embodiments, MenopauseComposition Formula Z is formulated as a gel.

Menopause Composition Formula AB comprises: 96.635% by weight purifiedwater (range 92%-98%); 1.5% by weight hypromellose (range 0.5%-2%);0.24% by weight carbomer (range 0.1%-1%); 0.7% by weight monobasicsodium phosphate (range 0.5%-1.0%); 0.165% by weight dibasic sodiumphosphate (range 0.01%-0.5%); optionally 0.232% by weight phenethylalcohol (range 0.1%-1% by weight); 0.15% by weight sodium chloride(range 0.01%-1%); optionally 0.168% by weight caprylyl glycol (range0.1%-1% by weight); 0.05% by weight Salvia sclarea (range 0.015%-2.5%);0.04% by weight xylose (range 0.015%-2.5%); and 0.12% by weight sodiumhydroxide (range 0.001%-1%). In certain embodiments, MenopauseComposition Formula AB is formulated as a gel.

In certain embodiments, a topical, isotonic, biome-friendly menopausecomposition of the present disclosure comprise xylose at a concentrationranging from about 0.01% to about 2.5% by weight and Salvia extract at aconcentration ranging from about 0.01% to about 2.5% by weight, andwherein the composition has a pH ranging from about 4.0 to about 6.8 andis optionally formulated as a gel. In some embodiments, the compositioncomprises xylose at a concentration ranging from about 0.02% to about0.5% by weight and Salvia extract at a concentration ranging from about0.01% to about 0.2% by weight. In some embodiments, the Salvia extractis Salvia sclarea, Salvia hispanica, Salvia plebeia, or combinationsthereof.

The menopause compositions may further comprise a non-irritatingviscosity increasing agent at a concentration ranging from about 0.1% toabout 10% by weight. In certain embodiments, the non-irritatingviscosity-increasing agent is a cellulose ester (e.g., methylcellulose,ethylcellulose, ethyl methyl cellulose, hydroxyethyl cellulose,hydroxypropyl cellulose, hydroxyethyl methyl cellulose, hyroxypropylmethyl cellulose (hypromellose), ethyl hydroxyethyl cellulose,carboxymethyl cellulose) carbomer, polyoxazoline, or any combinationthereof. In some embodiments, the non-irritating viscosity-increasingagent is hypromellose at a total concentration ranging from about 0.3%to about 4% by weight. In other embodiments, the non-irritatingviscosity-increasing agent is a carbomer at a total concentrationranging from about 0.1% to about 1% by weight. In yet other embodiments,the non-irritating viscosity-increasing agent is hypromellose andcarbomer homopolymer type B together at a total concentration rangingfrom about 0.1% to about 5% by weight. In yet further embodiments, theratio of hypromellose to carbomer homopolymer type B in the compositionranges from about 2:1 to about 8:1. In other embodiments, thenon-irritating viscosity-increasing agent is hypromellose andcarboxymethylcellulose together at a total concentration ranging fromabout 1% to about 3% by weight. In yet other embodiments, thenon-irritating viscosity-increasing agent is poly(2-methyl-2-oxazoline)at a total concentration ranging from about 1% to about 10% by weight.

The menopause compositions may also comprise a surfactant at a totalconcentration ranging from about 0.5% to about 5% by weight. In certainembodiments, the surfactant is a cetyl hydroxyethylcellulose, polyvinylalcohol, poloxamer, or any combination thereof. In some embodiments, thesurfactant is a cetyl hydroxyethylcellulose at a total concentrationranging from about 0.5% to about 2% by weight. In other embodiments, thesurfactant is a polyvinyl alcohol at a total concentration ranging fromabout 1% to about 2% by weight. In yet other embodiments, the surfactantis a poloxamer at a total concentration ranging from about 0.5% to about1.5% by weight.

The menopause compositions may further comprise a paraben-freepreservative at a total concentration ranging from about 0.005% to about5% by weight. In certain embodiments, the paraben-free preservative is aphenethyl alcohol, caprylyl glycol, phenoxyethanol, benzoic acid, abenzoate, a sorbate, cranberry extract, oleuropein, acemannan,carvacrol, or any combination thereof. In some embodiments, wherein theparaben-free preservative is a phenethyl alcohol and caprylyl glycoltogether at a total concentration ranging from about 0.25% to about 1.2%by weight. In other embodiments, the paraben-free preservative is aphenoxyethanol and caprylyl glycol at a total concentration ranging fromabout 0.25% to about 0.75% by weight. In yet other embodiments, theparaben-free preservative is benzoic acid at a total concentrationranging from about 0.025% to about 0.75% by weight. In yet otherembodiments, the paraben-free preservative is a potassium sorbate andsodium benzoate at a total concentration ranging from about 0.1% toabout 0.5% by weight.

The menopause compositions may further comprise a buffering agent at atotal concentration ranging from about 0.5% to about 4% by weight. Incertain embodiments, the buffering agent is a sodium phosphate(monobasic, dibasic, or both), potassium phosphate (monobasic, dibasic,or both), citric acid, sodium citrate, or any combination thereof. Insome embodiments, the buffering agent is a dibasic sodium phosphate anda monobasic sodium phosphate at a total concentration ranging from about0.5% to about 1.5% by weight. In yet other embodiments, the bufferingagent is citric acid and sodium citrate at a total concentration rangingfrom about 0.75% to about 1.5% by weight. In yet other embodiments, thebuffering agent is sodium phosphate at a total concentration rangingfrom about 0.25% to about 1.25% by weight.

The menopause compositions may further comprise an osmolality adjuster,such as sodium chloride, potassium chloride, or both, at a totalconcentration ranging from about 0.1% to about 1% by weight.

The menopause compositions may further comprise at least one pHmodifying agent at a total concentration ranging from about 0.001% toabout 2% by weight. At least one pH modifying agent may be an acidifyingagent, an alkalizing agent, or both. In some embodiments, the alkalizingagent is ammonia, ammonium carbonate, diethanolamine, monoethanolamine,potassium hydroxide, potassium phosphate dibasic, sodium bicarbonate,sodium borate, sodium carbonate, sodium hydroxide, sodium lactate,sodium phosphate dibasic, trolamine, or any combination thereof. In aspecific embodiment, the alkalizing agent is sodium hydroxide at a totalconcentration ranging from about 0.01% to about 1% by weight.

The menopause compositions may also include a solvent, such as water inan amount from about 85% to about 98%.

An exemplary topical, isotonic, biome-friendly menopause compositioncomprises xylose at a concentration ranging from about 0.01% to about2.5% by weight or about 0.02% to about 0.5% by weight and Salvia extractof the composition is Salvia sclarea at a concentration ranging fromabout 0.025% to about 0.075% by weight, and the composition furthercomprises: a non-irritating viscosity-increasing agent, wherein thenon-irritating viscosity-increasing agent is hypromellose at a totalconcentration ranging from about 0.75% to about 1.25% by weight; asurfactant, wherein the surfactant is polyvinyl alcohol and poloxamertogether at a total concentration ranging from about 2% to about 2.5% byweight; a buffering agent, wherein the buffering agent is a dibasicsodium phosphate and a monobasic sodium phosphate at a totalconcentration ranging from about 0.5% to about 1% by weight; anosmolality adjuster, wherein the osmolality adjuster is sodium chlorideat a total concentration ranging from about 0.1% to about 0.7% byweight; a solvent, wherein the solvent is water at a concentrationranging from about 95% to about 98% by weight; a pH modifier, whereinthe pH modifier is sodium hydroxide at a concentration ranging fromabout 0.001% to about 1% by weight; and optionally, a paraben-freepreservative, wherein the paraben-free preservative is preferablybenzoic acid at a total concentration ranging from about 0.05% to about0.25% by weight, and wherein the composition has a pH ranging from about4.5 to about 6.8. The menopause composition can be formulated as a gel.

Another exemplary topical, isotonic, biome-friendly menopausecomposition comprises xylose at a concentration ranging from about 0.01%to about 2.5% by weight or about 0.02% to about 0.5% by weight andSalvia sclarea at a concentration ranging from about 0.025% to about0.075% by weight, and the composition further comprises: anon-irritating viscosity-increasing agent, wherein the non-irritatingviscosity-increasing agent is hypromellose and carbomer homopolymer typeB together at a total concentration ranging from about 1.0% to about 2%by weight, and wherein the ratio of hypromellose to carbomer homopolymertype B in the composition is about 6.2:1; a buffering agent, wherein thebuffering agent is a dibasic sodium phosphate and a monobasic sodiumphosphate at a total concentration ranging from about 0.5% to about 1%by weight; an osmolality adjuster, wherein the osmolality adjuster issodium chloride at a total concentration ranging from about 0.1% toabout 0.7% by weight; a solvent, wherein the solvent is water at aconcentration ranging from about 92% to about 98% by weight; a pHmodifier, wherein the pH modifier is sodium hydroxide at a concentrationranging from about 0.001% to about 1% by weight; and optionally, aparaben-free preservative, wherein the paraben-free preservative ispreferably a phenethyl alcohol and caprylyl glycol together at a totalconcentration ranging from about 0.2% to about 0.75% by weight, andwherein the composition has a pH ranging from about 4.5 to about 6.8.The menopause composition can be formulated as a gel.

Another example of a topical, isotonic, biome-friendly menopausecomposition comprises xylose at a concentration ranging from about 0.01%to about 2.5% by weight or about 0.02% to about 0.5% by weight andSalvia sclarea at a concentration ranging from about 0.025% to about0.075% by weight, and the composition further comprises: anon-irritating viscosity-increasing agent, wherein the non-irritatingviscosity-increasing agent is hypromellose at a total concentrationranging from about 1.25% to about 2.25% by weight; a surfactant, whereinthe surfactant is poloxamer at a total concentration ranging from about0.75% to about 1.25% by weight; a buffering agent, wherein the bufferingagent is monobasic sodium phosphate and dibasic sodium phosphate at atotal concentration ranging from about 0.5% to about 1% by weight; anosmolality adjuster, wherein the osmolality adjuster is sodium chlorideat a total concentration ranging from about 0.1% to about 0.7% byweight; a solvent, wherein the solvent is water at a concentrationranging from about 95% to about 98% by weight; and optionally, aparaben-free preservative, wherein the paraben-free preservative ispreferably a cranberry extract at a total concentration ranging fromabout 0.01% to about 0.05% by weight, and wherein the composition has apH ranging from about 4.5 to about 6.8. The menopause composition can beformulated as a gel.

Yet another example of a topical, isotonic, biome-friendly menopausecomposition comprises xylose at a concentration ranging from about 0.01%to about 2.5% by weight or about 0.02% to about 0.5% by weight andSalvia plebeia at a concentration ranging from about 0.025% to about0.075% by weight, and the composition further comprises: anon-irritating viscosity-increasing agent, wherein the non-irritatingviscosity-increasing agent is carbomer homopolymer type B andhypromellose at a total concentration ranging from about 0.10% to about1% by weight, and wherein the ratio of hypromellose to carbomer in thecomposition is about 3:1; a buffering agent, wherein the buffering agentis monobasic sodium phosphate and dibasic sodium phosphate at a totalconcentration ranging from about 0.5% to about 1% by weight; anosmolality adjuster, wherein the osmolality adjuster is sodium chlorideat a total concentration ranging from about 0.1% to about 0.7% byweight; a pH modifier, wherein the pH modifier is sodium hydroxide at aconcentration ranging from about 0.001% to about 1%; a solvent, whereinthe solvent is water at a concentration ranging from about 95% to about99% by weight; optionally, a paraben-free preservative, wherein theparaben-free preservative is preferably phenethyl alcohol and caprylylglycol together at a total concentration ranging from about 0.25% toabout 0.5% by weight, and wherein the composition has a pH ranging fromabout 4.5 to about 6.8. The menopause composition can be formulated as agel.

Yet another exemplary topical, isotonic, biome-friendly menopausecomposition comprises xylose at a concentration ranging from about 0.01%to about 2.5% by weight or about 0.02% to about 0.5% by weight andSalvia sclarea at a concentration ranging from about 0.025% to about0.075% by weight, and the composition further comprises: anon-irritating viscosity-increasing agent, wherein the non-irritatingviscosity-increasing agent is hypromellose and carboxymethyl cellulosetogether at a total concentration ranging from about 1.5% to about 2.5%by weight, and wherein the ratio of hypromellose to carboxymethylcellulose in the composition is about 1.3:1; a buffering agent, whereinthe buffering agent is citric acid and sodium citrate together at atotal concentration ranging from about 1% to about 1.25% by weight; anosmolality adjuster, wherein the osmolality adjuster is sodium chlorideat a total concentration ranging from about 0.1% to about 0.7% byweight; a solvent, wherein the solvent is water at a concentrationranging from about 95% to about 98% by weight; a pH modifier, whereinthe pH modifier is sodium hydroxide at a concentration ranging fromabout 0.001% to about 1%; and optionally, a paraben-free preservative,wherein the paraben-free preservative is preferably potassium sorbateand sodium benzoate together at a total concentration ranging from about0.25% to about 0.5% by weight, wherein the composition has a pH rangingfrom about 4.5 to about 6.8. The menopause composition can be formulatedas a gel.

(5) Freshening Compositions

In yet another aspect of the present disclosure, freshening compositionsfor the female lower reproductive tract are provided. Douching iscommonly performed by women seeking cleanliness of LRT and control ofunwanted feminine odors. These odors arise following vaginal infection,sweating in the external genital area, following sexual intercourse, andfollowing menses. However, douching is associated with dramaticincreases in LRT cancer, STD infection, bacterial vaginosis,infertility, and abnormal birth outcomes. Use of low pH “balancing” gelsthat bind to the vaginal wall and provide long-lasting effect has beenshown to significantly lower vaginal pH and bacterial vaginosissymptoms. However, gels that are currently sold for this use causemoderate to severe LRT cell toxicity and damage to vaginalLactobacillus, which are requisite for vaginal homeostasis. Thefreshening compositions of the present disclosure can be used to freshenor cleanse LRT (e.g., vagina or vulva), control vaginal odor, and are pHmatched to the vulvar and vaginal pH of the subject, e.g., for areproductive aged woman (about 3.5 to about 5.5), an ovulatory phasewoman (about 6.8 to about 7.8), pregnant woman (about pH 4.5 to about6.5), or postmenopausal woman (around about 4.5 to about 6.8), arenon-irritating for the LRT, and support homeostasis of the LRT,including preserving vaginal microbiota.

Freshening compositions of the present disclosure can be formulated as asemi-solid, gel, ointment, liquid, lotion, jelly, film, suspension,emulsion, mucilage, foam, cream, paste, or aerosol.

In some embodiments the freshening compositions are formulated as abioadhesive gel or film, so that they will not leak out of the vaginafollowing application and can provide pH adjustment to the LRT mucosafor 24 hours or more.

Freshening compositions of the present disclosure (e.g., FresheningComposition Formulas W, X, Y, Z, AB, AZ, BY) can be used for perinealmassage, contraception, anti-microbial, fertility enhancement, sexuallubrication, vaginal hydration, sexual arousal, vaginal tightening,vaginal freshening, vaginal homeostasis, as a drug carrier base, or anycombination thereof.

In certain embodiments, the freshening compositions may be used inmethods of cleansing the lower reproductive tract or controllingunwanted feminine odor, comprising administering the fresheningcomposition to the lower reproductive tract of a female subject (e.g.,vagina or vulva).

Exemplary formulations of freshening compositions and % ranges of eachcomponent as contemplated in the present disclosure are set forth asfollows.

Freshening Composition Formula W comprises: 90.89% by weight purifiedwater (range 50%-99%); 2.0% by weight polyvinyl alcohol (range0.01%-5%); 4.8% by weight cetyl hydroxyethyl cellulose (range 0.1%40%),1.0% by weight poly(2-methyl-2-oxazoline) (range 0.5%-40%); 0.139% byweight lactic acid (range 0.01%-2%); 0.683% by weight sodium lactate(range 0.01%-5%); 0.05% by weight Salvia sclarea (range 0.01%-1%); 0.2%by weight sodium chloride (range 0.01%-1%); 0.04% by weight xylose(range 0.01%-5%), and 0.12% by weight sodium hydroxide (range0.001%-1%). In certain embodiments, Freshening Composition Formula W isformulated as a gel.

Freshening composition formula X comprises: 89.45% by weight purifiedwater (range 50%-99%); 9.0% by weight poly(2-methyl-2-oxazoline) (range0.5%-40%); 0.139% by weight lactic acid (range 0.01%-2%); 0.683% byweight sodium lactate (range 0.01%-5%); 0.232% by weight phenethylalcohol (range 0.01%-1%); 0.168 caprylyl glycol (0.01%-2%); 0.05% byweight Salvia sclarea (range 0.01%-1%); 0.2% by weight sodium chloride(range 0.01%-1%); 0.04% by weight xylose (range 0.01%-5%); and 0.04% byweight (range 0.001%-1%). In certain embodiments, Freshening CompositionFormula X is formulated as a gel. Freshening composition formula Ycomprises: 95.16% by weight purified water (range 50%-99%); 1.0% byweight cetyl hydroxyethylcellulose (range 0.1%-10%); 1.5% by weightcarbomer (range 0.01%-5%); 0.66% by weight ascorbic acid (range0.01%-5%); 0.49% by weight sodium ascorbate (range 0.01%-5%); 0.232% byweight phenethyl alcohol (range 0.01%-1%); 0.168% by weight caprylylglycol (range 0.01%-2%); 0.05% by weight Salvia sclarea (range0.01%-1%); 0.3% by weight sodium chloride (range 0.01%-1%); and 0.08% byweight xylose (range 0.01%-5%). In certain embodiments, FresheningComposition Formula Y is formulated as a gel.

Freshening composition formula Z comprises: 95.80% by weight purifiedwater (range 50%-99%); 2.0% by weight polyvinyl alcohol (range0.01%-5%); 1.0% by weight carbomer (0.01%-5%); 0.139% by weight lacticacid (range 0.01%-2%); 0.683% by weight sodium lactate (range 0.01%-5%);0.05% by weight Salvia sclarea (range 0.01%-1%); 0.2% by weight sodiumchloride (range 0.01%-1%); 0.04% by weight xylose (range 0.01%-5%); and0.009% by weight sodium hydroxide (range 0.001%-1%). In certainembodiments, Freshening Composition Formula Z is formulated as a gel.

Freshening composition formula AZ comprises: 83.9% by weight purifiedwater (range 50%-99%); 0.1% by weight Salvia plebeia (range 0.01%-1%);0.03% by weight cranberry extract (0.01%-5%); 0.5% by weight sodiumchloride (0.01%-1%); 0.04% by weight xylose (0.01%-5%); 15% by weightpoly(2-methyl-2-oxazoline) (range 0.5%-40%); and 0.43% by weight sodiumhydroxide (range 0.001%-1%). In certain embodiments, FresheningComposition Formula AZ is formulated as a gel.

Freshening composition formula BY comprises: 96.03% by weight purifiedwater (range 50%-99%); 0.04% by weight xylose (0.01%-5%); 0.4% by weightmonobasic sodium phosphate (range 0.01%-2%); 0.7% by weight sodiumcitrate; 0.25% by weight caprylyl glycol (range 0.01%-2%); 0.14% byweight sodium chloride (range 0.01%-1%); 1.1% by weight hypromellose(range 0.1%-10%); 0.24% by weight carbomer (range 0.01%-5%); 0.1% byweight Salvia plebeia (range 0.01%-1%); and 1.0% by weight polyvinylalcohol (range in a quantity required to complete 100%). In certainembodiments, Freshening Composition Formula BY is formulated as a gel.

Freshening composition formula AB comprises: 95.328% by weight purifiedwater (range 90%-99%); 1.0% by weight cetyl hydroxyethylcellulose (range0.25%-10%); 2.0% by weight carbomer (range 0.1%-3%); 0.139% by weightlactic acid (range 0.01%-0.5%); 0.683% by weight sodium lactate (range0.05%-1%); 0.05% by weight Salvia sclarea (range 0.025%-2.5%); 0.2% byweight sodium chloride (range 0.1%-1%); 0.04% by weight xylose (range0.025%-2.5%); 0.16% by weight sodium hydroxide (range 0.001%-1%);optionally 0.232% by weight phenethyl alcohol (range 0.1%-1%); andoptionally 0.168% by weight caprylyl glycol (range 0.1%-1%).

In certain embodiments, topical, isotonic, biome-friendly fresheningcompositions of the present disclosure comprise xylose at aconcentration ranging from about 0.01% to about 2.5% by weight and aSalvia extract at a concentration ranging from about 0.01% to about 2.5%by weight, and wherein the composition has a pH ranging from about 3.5to about 6.8 and is optionally formulated as a gel. The pH of thefreshening compositions may be matched to the life-specific of menstrualcycle specific stage of the subject, e.g., about pH 4.5 to about 6.8 formenopausal female subjects; about pH 3.5 to about 5.5 for reproductiveaged female subjects that are not in the ovulatory phase; or about pH4.5 to about 6.5 for pregnant subjects. In some embodiments, xylose isin an amount ranging from about 0.02% to about 0.5% by weight and Salviaextract is at a concentration ranging from about 0.01% to about 0.2% byweight. In some embodiments, the Salvia extract is Salvia sclarea,Salvia hispanica, Salvia plebeia, or combinations thereof.

The freshening compositions may further comprise a non-irritatingviscosity increasing agent at a concentration ranging from about 0.1% toabout 25% by weight. In certain embodiments, the non-irritatingviscosity-increasing agent is a cellulose ester (e.g., methylcellulose,ethylcellulose, ethyl methyl cellulose, hydroxyethyl cellulose,hydroxypropyl cellulose, hydroxyethyl methyl cellulose, hyroxypropylmethyl cellulose (hypromellose), ethyl hydroxyethyl cellulose,carboxymethyl cellulose), carbomer, polyoxazoline, or any combinationthereof. In some embodiments, the non-irritating viscosity-increasingagent is a carbomer, such as homopolymer type B at a total concentrationranging from about 0.1% to about 3% by weight. In some embodiments, thenon-irritating viscosity-increasing agent is a polyoxazoline, such aspoly(2-methyl-2-oxazoline) at a total concentration ranging from about5% to about 10%.

The freshening compositions may further comprise a surfactant at a totalconcentration ranging from about 0.25% to about 10% by weight. Incertain embodiments, the surfactant is cetyl hydroxyethylcellulose,polyvinyl alcohol, or both. In some embodiments, the surfactant is acetyl hydroxyethylcellulose at a total concentration ranging from about0.25% to about 5% by weight. In other embodiments, the surfactant is apolyvinyl alcohol at a total concentration ranging from about 1% toabout 3% by weight.

The freshening compositions may also include a paraben-free preservativeat a total concentration ranging from about 0.005% to 5% by weight. Incertain embodiments, the paraben-free preservative is a phenethylalcohol, caprylyl glycol, oleuropein, acemannan, carvacrol, anascorbate, cranberry extract, or any combination thereof. In someembodiments, the paraben-free preservative is a phenethyl alcohol andcaprylyl glycol together at a total concentration ranging from about0.02% to about 1.5% by weight. In other embodiments, the paraben-freepreservative is a phenethyl alcohol, caprylyl glycol and a sodiumascorbate, together at a total concentration ranging from about 0.5% toabout 2% by weight. In yet other embodiments, the paraben-freepreservative is a cranberry extract at a total concentration rangingfrom about 0.01% to about 2.5% by weight.

The freshening compositions may further comprise a buffering agent at atotal concentration ranging from about 0.05% to about 2.5% by weight. Incertain embodiments, the buffering agent is lactic acid, sodium lactate,ascorbic acid, sodium ascorbate, sodium phosphate, acetic acid, sodiumacetate, citric acid, or any combination thereof. In some embodiments,the buffering agent is lactic acid and sodium lactate together at atotal concentration ranging from about 0.4% to about 1% by weight. Inother embodiments, the buffering agent is ascorbic acid and sodiumascorbate at a total concentration ranging from about 0.5% to about 2%by weight.

The freshening compositions may also comprise an osmolality adjuster,such as sodium chloride, potassium chloride, or both, at a totalconcentration ranging from about 0.1% to about 1% by weight.

The freshening compositions may further comprise at least one pHmodifying agent, such as an acidifying agent, an alkalizing agent, orboth, at a total concentration ranging from about 0.001% to about 2% byweight. In some embodiments, the alkalizing agent is ammonia, ammoniumcarbonate, diethanolamine, monoethanolamine, potassium hydroxide,potassium phosphate dibasic, sodium bicarbonate, sodium borate, sodiumcarbonate, sodium hydroxide, sodium lactate, sodium phosphate dibasic,trolamine, or any combination thereof. In a specific embodiment, thealkalizing agent is sodium hydroxide at a total concentration rangingfrom about 0.01% to about 1% by weight.

The freshening compositions may also comprise a solvent. In a specificembodiment, the solvent is water and the composition comprises fromabout 90% to about 99% water.

An exemplary topical, isotonic, biome-friendly freshening compositioncomprises xylose at a concentration ranging from about 0.01% to about2.5% by weight or about 0.02% to about 0.5% by weight and Salvia sclareaat a concentration ranging from about 0.025% to about 0.075% by weight,and the composition further comprises: a non-irritatingviscosity-increasing agent, wherein the non-irritatingviscosity-increasing agent is poly(2-methyl-2-oxazoline) at a totalconcentration ranging from about 1% to about 10% by weight; asurfactant, wherein the surfactant is polyvinyl alcohol and cetylhydroxyethylcellulose together at a total concentration ranging fromabout 1% to about 9% by weight; a buffering agent, wherein the bufferingagent is lactic acid and sodium lactate together at a totalconcentration ranging from about 0.5% to about 1% by weight; anosmolality adjuster, wherein the osmolality adjuster is sodium chlorideat a total concentration ranging from about 0.1% to about 0.7% byweight; a pH modifier, wherein the pH modifier is sodium hydroxide at aconcentration ranging from about 0.001% to about 1% by weight; and asolvent, wherein the solvent is water at a concentration ranging fromabout 90% to about 98% by weight, and wherein the composition has a pHranging from about 3.5 to about 6.8. The freshening composition can beformulated as a gel. Another exemplary topical, isotonic, biome-friendlyfreshening composition comprises xylose at a concentration ranging fromabout 0.01% to about 2.5% by weight or about 0.02% to about 0.5% byweight and Salvia sclarea at a concentration ranging from about 0.025%to about 0.075% by weight, and the composition further comprises: anon-irritating viscosity-increasing agent, wherein the non-irritatingviscosity-increasing agent is poly(2-methyl-2-oxazoline) at a totalconcentration ranging from about 5% to about 10% by weight; a bufferingagent, wherein the buffering agent is lactic acid and sodium lactatetogether at a total concentration ranging from about 0.5% to about 1% byweight; an osmolality adjuster, wherein the osmolality adjuster issodium chloride at a total concentration ranging from about 0.1% toabout 0.7% by weight; a solvent, wherein the solvent is water at aconcentration ranging from about 88% to about 98% by weight; a pHmodifier, wherein the pH modifier is sodium hydroxide at a concentrationranging from about 0.001% to about 1%; and optionally, a paraben-freepreservative, wherein the paraben-free preservative is preferably aphenethyl alcohol and caprylyl glycol together at a total concentrationranging from about 0.2% to about 0.7% by weight, and wherein thecomposition has a pH ranging from about 3.5 to about 6.8. The fresheningcomposition can be formulated as a gel.

Yet another example of a topical, isotonic, biome-friendly fresheningcomposition comprises xylose at a concentration ranging from about 0.01%to about 2.5% by weight or about 0.02% to about 0.5% by weight andSalvia sclarea at a concentration ranging from about 0.025% to about0.075% by weight, and the composition further comprises: anon-irritating viscosity-increasing agent, wherein the non-irritatingviscosity-increasing agent is carbomer homopolymer type B at a totalconcentration ranging from about 0.2% to about 3% by weight; asurfactant, wherein the surfactant is cetyl hydroxyethylcellulose at atotal concentration ranging from about 0.5% to about 1.5% by weight; abuffering agent, wherein the buffering agent is lactic acid and sodiumlactate together at a total concentration ranging from about 0.4% toabout 1% by weight; an osmolality adjuster, wherein the osmolalityadjuster is sodium chloride at a total concentration ranging from about0.1% to about 0.7% by weight; a solvent, wherein the solvent is water ata concentration ranging from about 90% to about 98% by weight; a pHmodifier, wherein the pH modifier is sodium hydroxide at a concentrationranging from about 0.001% to about 1% by weight; and optionally, aparaben-free preservative, wherein the paraben-free preservative ispreferably a phenethyl alcohol and caprylyl glycol together at a totalconcentration ranging from about 0.2% to about 0.7% by weight, andwherein the composition has a pH ranging from about 3.5 to about 6.8.The freshening composition can be formulated as a gel.

Another exemplary topical, isotonic, biome-friendly fresheningcomposition comprises xylose at a concentration ranging from about 0.01%to about 2.5% by weight or about 0.02% to about 0.5% by weight andSalvia sclarea at a concentration ranging from about 0.025% to about0.075% by weight, and the composition further comprises: anon-irritating viscosity-increasing agent, wherein the non-irritatingviscosity-increasing agent is carbomer homopolymer type B at a totalconcentration ranging from about 0.1% to about 3% by weight; asurfactant, wherein the surfactant is cetyl hydroxyethylcellulose at atotal concentration ranging from about 0.5% to about 1.5% by weight; abuffering agent, wherein the buffering agent is ascorbic acid and sodiumascorbate together at a total concentration ranging from about 0.5% toabout 1.2% by weight; an osmolality adjuster, wherein the osmolalityadjuster is sodium chloride at a total concentration ranging from about0.1% to about 0.7% by weight; a solvent, wherein the solvent is water ata concentration ranging from about 95% to about 98% by weight; andoptionally, a paraben-free preservative, wherein the paraben-freepreservative is preferably a phenethyl alcohol and caprylyl glycoltogether at a total concentration ranging from about 0.2% to about 0.7%by weight, and wherein the composition has a pH ranging from about 3.5to about 6.8. The freshening composition can be formulated as a gel.

Yet another exemplary topical, isotonic, biome-friendly fresheningcomposition comprises xylose at a concentration ranging from about 0.01%to about 2.5% by weight or about 0.02% to about 0.5% by weight andSalvia sclarea at a concentration ranging from about 0.025% to about0.075% by weight, and the composition further comprises: anon-irritating viscosity-increasing agent, wherein the non-irritatingviscosity-increasing agent is carbomer homopolymer type B at a totalconcentration ranging from about 1% to about 3% by weight; a surfactant,wherein the surfactant is polyvinyl alcohol at a total concentrationranging from about 1% to about 3% by weight; a buffering agent, whereinthe buffering agent is lactic acid and sodium lactate together at atotal concentration ranging from about 0.5% to about 1% by weight; anosmolality adjuster, wherein the osmolality adjuster is sodium chlorideat a total concentration ranging from about 0.1% to about 0.7% byweight; a solvent, wherein the solvent is water at a concentrationranging from about 95% to about 98% by weight; and a pH modifier,wherein the pH modifier is sodium hydroxide at a concentration rangingfrom about 0.001% to about 1% by weight, and wherein the composition hasa pH ranging from about 3.5 to about 6.8. The freshening composition canbe formulated as a gel.

Another exemplary topical, isotonic, biome-friendly fresheningcomposition comprises xylose at a concentration ranging from about 0.01%to about 2.5% by weight or about 0.02% to about 0.5% by weight andSalvia plebeia at a concentration ranging from about 0.025% to about0.25% by weight, and the composition further comprises: a non-irritatingviscosity-increasing agent, wherein the non-irritatingviscosity-increasing agent is poly(2-methyl-2-oxazoline) at a totalconcentration ranging from about 5% to about 17% by weight; anosmolality adjuster, wherein the osmolality adjuster is sodium chlorideat a total concentration ranging from about 0.1% to about 0.7% byweight; a solvent, wherein the solvent is water at a concentrationranging from about 80% to about 95% by weight; and a pH modifier,wherein the pH modifier is sodium hydroxide at a concentration rangingfrom about 0.001% to about 1% by weight, and optionally, a paraben-freepreservative, wherein the paraben-free preservative is preferablycranberry extract at a total concentration from about 0.01% to about0.5%, wherein the composition has a pH ranging from about 3.5 to about6.8. The freshening composition can be formulated as a gel.

Another exemplary topical, isotonic, biome-friendly fresheningcomposition comprises xylose at a concentration ranging from about 0.01%to about 2.5% by weight or about 0.02% to about 0.5% by weight andSalvia plebeia at a concentration ranging from about 0.025% to about0.25% by weight, and the composition further comprises: a non-irritatingviscosity-increasing agent, wherein the non-irritatingviscosity-increasing agent is hypromellose and carbomer homopolymer typeB together at a total concentration ranging from about 1.0% to about 2%by weight, and wherein the ratio of hypromellose to carbomer homopolymertype B in the composition is about 4.5:1; a surfactant, wherein thesurfactant is polyvinyl alcohol at a total concentration ranging fromabout 0.8% to about 3% by weight; a buffering agent, wherein thebuffering agent is monobasic sodium phosphate and sodium citrate at atotal concentration ranging from about 0.5% to about 1.5% by weight; anosmolality adjuster, wherein the osmolality adjuster is sodium chlorideat a total concentration ranging from about 0.1% to about 0.7% byweight; a solvent, wherein the solvent is water at a concentrationranging from about 95% to about 98% by weight; and optionally, aparaben-free preservative, wherein the paraben-free preservative ispreferably caprylyl glycol at a total concentration from about 0.1% toabout 0.5%, wherein the composition has a pH ranging from about 3.5 toabout 6.8. The freshening composition can be formulated as a gel.

(6) Vaginal Tightening Compositions

In yet another aspect of the present disclosure, vaginal tighteningcompositions are provided. Compositions (e.g., gels) that create asensation of greater friction during intercourse are desired by manywomen and couples, especially after childbirth and aging. In addition,several cultures perceive a benefit of “dry sex” with minimallubrication of the vaginal canal. Common ingredients in availableproducts for tightening the vagina contain highly drying and irritatingcompounds, including talc, alum, and witch hazel. Many women “wash” thevagina out prior to intercourse with water and washcloths to reducenormal arousal fluids and perceived “excess” lubrication. Thesecompounds and practices dry and increase microtrauma in the vaginalvault, which can increase STD transmission risks. The vaginal tighteningcompositions (e.g., bioadhesive vaginal gels or films) provided hereincreate a sense of greater friction in the vagina. The vaginal tighteningcompositions coat and protect the vagina, but also allow for thesensation of friction during intercourse.

The vaginal tightening compositions are pH matched to the vulvar andvaginal pH of the subject, e.g., for a reproductive aged woman (about pH3.5 to about 5.5), an ovulatory phase woman (about 6.8 to about 7.8),pregnant woman (about pH 4.5 to about 6.5), or postmenopausal woman(around about 4.0 to about 6.8). In certain embodiments, the vaginaltightening compositions comprise a poly(2-oxazoline). Thus, for example,any of the LRT compositions disclosed herein that further comprisepoly(2-oxazoline) may be used as vaginal tightening compositions. Apoly(2-oxazoline) may be poly(2-alkyl-2-oxazoline) orpoly(2-aryl-2-oxazoline), or both. In some embodiments, apoly(2-alkyl-2-oxazoline) may be poly(2-methyl-2-oxazoline),poly(2-ethyl-2-oxazoline), poly(2-isopropyl-2-oxazoline), or anycombination thereof. In some embodiments, a poly(2-aryl-2-oxazoline) ispoly(2-phenyl-2-oxazoline). Poly(2-oxazoline) is a low viscosity,hydrophilic, biocompatible polymer that provides firm bioadhesion to thevaginal wall, thereby protecting the LRT from micro trauma, while at thesame time providing friction of the penis against the bioadhesivecoating.

Vaginal tightening compositions of the present disclosure can beformulated as a semi-solid, gel, ointment, liquid, lotion, jelly, film,suspension, emulsion, mucilage, foam, cream, paste, or aerosol.

Vaginal tightening compositions of the present disclosure can be usedfor perineal massage, contraception, anti-microbial, fertilityenhancement, sexual lubrication, vaginal hydration, sexual arousal,vaginal tightening, vaginal freshening, vaginal homeostasis, as a drugcarrier base, or any combination thereof.

In certain embodiments, the vaginal tightening compositions are used inmethods for increasing friction during sexual intercourse, comprisingadministering the vaginal tightening composition intravaginally, to thepenis, to condoms, or to other devices (e.g., sex toys). In someembodiments, the vaginal tightening composition is administered within24 hours, 12 hours, 6 hours, 2 hours, 1 hour, 30 minutes, 15 minutes, 5minutes, just prior to, or during sexual intercourse.

Exemplary formulations of vaginal tightening compositions and % rangesof each component as contemplated in the present disclosure are setforth as follows.

Vaginal tightening composition formula 1 comprises: 89.1% by weightpurified water; 0.1% by weight Salvia plebeia; 0.406% by weightphenethyl alcohol; 0.294% by weight caprylyl glycol; 0.5% by weightsodium chloride; 0.4% by weight xylose; 9.1% by weightpoly(2-methyl-oxazoline) 25 kDa; 0.01% by weight sodium hydroxide. Incertain embodiments, vaginal tightening composition formula A isformulated as a gel.

Vaginal tightening composition formula 2 comprises: 83.5% by weightpurified water; 0.1% by weight Salvia plebeia; 0.406% by weightphenethyl alcohol; 0.294% by weight caprylyl glycol; 0.5% by weightsodium chloride; 0.4% by weight xylose; 14.7% by weightpoly(2-ethyl-2-oxazoline) 70 kDa; 0.01% by weight sodium hydroxide. Incertain embodiments, vaginal tightening composition formula B isformulated as a gel.

(7) Drug Carrier Base Compositions

In yet another aspect of the present disclosure, the compositionsprovided herein (e.g., fertility compositions, contraceptivecompositions, perineal massage compositions, menopause compositions,freshening compositions) may be used as a drug carrier base. Forexample, the compositions may be combined with over the counter orprescription drugs to treat medical conditions of the female lowerreproductive tract. Common topical yeast treatments include 5 or 6inactive ingredients that compromise LRT homeostasis and function. Theirritation and burning that accompany these yeast infection treatmentscan actually lead to more LRT pathology and symptoms. The same is truefor hormone replacement therapy medications and a variety of other“therapies.” For example, a leading vaginal estrogen product containscornstarch as a primary ingredient, which causes irritation in manywomen, as well as an unsightly white discharge.

The drug carrier base compositions of the present disclosure preserveLRT homeostasis, and the “inactive” ingredients in the composition aretruly inactive. Drug carrier base compositions of the present disclosurecan be formulated as a semi-solid, gel, ointment, liquid, lotion, jelly,film, suspension, emulsion, mucilage, foam, cream, paste, or aerosol. Incertain embodiments, the drug carrier base compositions are formulatedas a bioadhesive formulation (e.g., gel or film) so that the medicationdose is retained inside the vagina, reducing loss of dosing and mess forfemale subject.

Drug carrier base compositions of the present disclosure can be used forperineal massage, contraception, anti-microbial, fertility enhancement,sexual lubrication, vaginal hydration, sexual arousal, vaginaltightening, vaginal freshening, vaginal homeostasis, as a drug carrierbase, or any combination thereof.

Examples of conditions that may be treated include yeast infections,bacterial infections, viral infections, hormone imbalances, infertility,and menopausal symptoms. In certain embodiments, the compositions of thepresent disclosure may be a drug carrier base for a topical painrelieving agent (topical analgesic), an anti-microbial (e.g.,antibiotic, anti-viral agent, anti-fungal agent, or a combinationthereof), an agent for treating or preventing a hormone imbalance, or anagent for treating or preventing infertility, or a combination thereof.

In certain embodiments, a topical pain relieving agent that is containedin a drug carrier base composition is lidocaine, benzocaine, novocaine,diphenhydramine, or pramoxine.

In certain embodiments, an anti-microbial agent is an anti-fungal agentselected from butoconazole nitrate, clotrimazole, miconazole nitrate,terconazole, tioconazole, econazole nitrate, efinaconazole,ketoconazole, luliconazole, naftifine hydrochloride, oxiconazolenitrate, sertaconazole nitrate, sulconazole nitrate, tavaborole,terbinafine, acyclovir, tenovir, zidovudine, stavudine, metronidazole,or a combination thereof.

Other examples of therapeutic agents include hormones (e.g., estradiol,estriol, estropipate, testosterone, progesterone, DHEA or a combinationthereof), contraceptive agents (e.g., that impair sperm function,thicken cervical mucus, or both), growth factors (e.g., vascularendothelial growth factor), and vasodilators (e.g., L-arginine, niacin,nicotinamide, alprostadil, phosphodiesterase inhibitor).

In certain embodiments, the compositions disclosed herein (e.g.,fertility compositions, contraceptive compositions, perineal massagecompositions, menopause compositions, freshening compositions) furthercomprise factors that maintain, improve, or enhance VMB (e.g.,Lactobacilli) function and health, such as amylase, glycogen, D-lacticacid, L-lactic acid, a prebiotic (e.g., acemannan or galactoarabinan),or a combination thereof.

In certain embodiments, the compositions disclosed herein furthercomprise at least one vaginal probiotic bacterial species or strain(e.g., Lactobacillus species).

B. In Vitro and In Vivo Activity

(1) Sperm Function Assays

Formulations and compositions of this disclosure can be evaluated fortheir effect on sperm survival and function through the use of assays ormodels known in the art, such as sperm motility assays (e.g., subjectiveor computer assisted), sperm viability studies, in vitro fertilizationand embryo development animal models, membrane integrity of sperm,survival time in culture, cervical mucus penetration, lipidperoxidation, capacitation, zona recognition, acrosome reaction andsperm-oocyte fusion, and sperm chromatin testing (reviewed in, e.g.,Vasan, Indian J. Urol. 27:41, 2011; Oehninger et al., Fertil. Steril.102:1528, 2014; Mortimer et al., 2013, Hum. Reprod. Update 19 (Suppl1):i1-i45). Additional testing can include post-coital testing toevaluate sperm presence in the cervical canal, and even pregnancyoutcomes in an animal model or among women in a clinical trial.

Sperm motility is one function that may be used to assess sperm functionand thus fertilization potential. Motility of sperm is expressed as thetotal percent of motile sperm, the total percent of progressively motilesperm (swimming forward), or the speed of sperm that are progressivelymotile. These measurements may be made by a variety of assays, but areconveniently assayed in one of two ways. Either a subjective visualdetermination is made using a phase contrast microscope when the spermare placed in a hemocytometer or on a microscope slide, or a computerassisted semen analyzer is used. Under phase contrast microscopy, motileand total sperm counts are made and speed is assessed as fast, medium orslow. A more specific of sperm motility is motility grade, where themotility of sperm is divided into four different grades (Cooper et al.,2010, Human Reprod. Update 16:231-45). Grade A refers to sperm withprogressive motility that are the strongest and swim fastest in astraight line. Grade B refers to sperm with non-linear motility; thatmove forward but tend to travel in a more curved or crooked motion.Grade C sperm have non-progressive motility in that they do not moveforward despite tail movement. Grade D sperm are immotile. Using acomputer assisted semen analyzer (such as IVOS Hamilton Thorne, Beverly,Mass.), the motility characteristics of individual sperm cells in asample are objectively determined (1998, Hum. Reprod. 13:142-5).Briefly, a sperm sample is placed onto a slide or chamber designed forthe analyzer. The analyzer tracks individual sperm cells and determinesmotility and velocity of the sperm. Data is expressed as percent motile,and measurements are obtained for path velocity and track speed as well.

Sperm viability can be measured using several different methods. By wayof example, two of these methods are staining with membrane exclusionstains and measurement of ATP levels. Briefly, a sample of sperm isincubated with a viable dye, such as Hoechst 33258 or eosin-nigrosinstain. Cells are placed in a hemocytometer and examined microscopically.Dead sperm with disrupted membranes stain with these dyes. The number ofcells that are unstained is divided by the total number of cells countedto give the percent live cells. ATP levels in a sperm sample aremeasured by lysing the sperm and incubating the lysate with luciferase,an enzyme obtained from fireflies, which fluoresces in the presence ofATP. The fluorescence is measured in a luminometer. The amount offluorescence in the sample is compared to the amount of fluorescence ina standard curve allowing a determination of the number of live spermpresent in the sample. Membrane integrity of sperm is typically assayedby a hypo-osmotic swell test which measures the ability of sperm to pumpwater or salts if exposed to non-isotonic environments. Briefly, in thehypo-osmotic swell test, sperm are suspended in a solution of 75 mMfructose and 25 mM sodium citrate, which is a hypo-osmotic (150 mOsm)solution. Sperm with intact, healthy membranes pump salt out of the cellcausing the membranes to shrink as the cell grows smaller. The spermtail curls inside this tighter membrane. Thus, sperm with curled tailare counted as live, healthy sperm with normal membranes. When comparedto the total number of sperm present, a percent of functional sperm maybe established.

The degree of membrane integrity is preferably determined by lipidperoxidation (LPO) measurements, which assess sperm membrane damagegenerated by free radicals released during handling. Lipid membraneperoxidation is assayed by incubating sperm with ferrous sulfate andascorbic acid for one hour in a 37° C. water bath. Proteins areprecipitated with ice-cold trichloroacetic acid. The supernatant iscollected by centrifugation and reacted by boiling with thiobarbituricacid and NaOH. The resultant malondialdehyde (MDA) formation isquantified by measuring absorbance at 534 nm as compared to an MDAstandard (Bell et al., J. Andrology 14:472-478, 1993). LPO is expressedas nM MDA/10⁸ sperm. A composition disclosed herein has a stabilizingeffect on sperm if exposure results in decreased LPO production.

The stability of chromatin DNA is assayed using the sperm chromatinstructure assay (SCSA). Sperm cells are very sensitive to oxidativestress resulting in sperm chromatin (DNA) damage (Whittington et al.,1999, Int. J. Andrology 22:236-242; Pasqualotto et al., 2000, Fertilityand Sterility 73:459-464; Kodama et al., 1997, Fertility and Sterility68:519-524). This damage can be profound in sperm cells because theycontain little to no mechanisms to repair DNA damage after it occurs.Substantial scientific data over the last decade has shown that spermchromatin damage can result in severe disruptions in fertility andadverse consequences for offspring (World Health Organization. WHOLaboratory Manual for the Examination and Processing of Human Semen.5^(th) ed. Switzerland: World Health Organization, 2010). This assay isbased on the metachromatic staining of single and double stranded DNA byacridine orange stain (Evenson et al., 1999, Human Reprod. 14:1039-1049;Evenson et al., 2002, J. Andrology 23:25-43). Excitation with an argonlaser causes acridine orange intercalated in double-stranded DNA to emita green fluorescence, whereas red fluorescence is emitted by singlestrand DNA. The extent of DNA denaturation in a sample is expressed as aand calculated by the formula α=red/(red+green). The endpointmeasurement is DNA Fragmentation Index (DFI). A DFI of <15% DFIindicates excellent to good sperm DNA integrity. Fresh sperm samples areincubated for a period of time in the presence of a test composition,flash frozen, and subsequently assayed for DNA breakage (see, e.g.,Evenson et al., 2002, J. Androl. 23:25-43).

In vitro fertilization rates are determined by measuring the percentfertilization of oocytes in vitro in an animal model such as bovine ormurine model. For example, maturing bovine oocytes are cultured in vitroin M199 medium plus 7.5% fetal calf serum and 50 μg/ml luteinizinghormone for 22 hours. Following culture for 4 hours, the sperm arechemically capacitated by adding 10 IU of heparin and incubated withbovine oocytes for 24 hours. At the end of the incubation, oocytes arestained with an aceto-orcein stain or equivalent to determine thepercent oocytes fertilized. Alternatively, fertilized oocytes may beleft in culture for 2 days, during which division occurs and the numberof cleaving embryos (i.e., 2 or more cells) is counted.

Survival time in culture of sperm (time to loss of motility) is anotherconvenient method of establishing sperm function. This parametercorrelates well with actual fertility of a given male. Briefly, analiquot of sperm is placed in culture medium, such as Tyrode's medium,pH 7.4 and incubated at 37° C., 5% CO₂, in a humidified atmosphere. Attimed intervals, for example every 2 hours, the percentage of motilesperm in the culture is determined by visual analysis using an invertedmicroscope or with a computer assisted sperm analyzer. As an endpoint, asperm sample is considered no longer viable when less than 5% of thecells have progressive motility.

Another parameter of sperm function is the ability to of sperm to swimup into a column of cervical mucus or substitute (reviewed in Ola etal., Hum. Reprod. 18:1037-1046, 2003). This cervical mucus penetrationtest can be done either in vitro or in vivo. Sperm are placed at one endof the track and the distance that sperm have penetrated into the mucusafter a given time period is determined. Cervical mucus penetrationstudies offer valuable biocompatibility data for devices that are usedfor reproductive purposes. The bovine cervical mucus penetration studyis an excellent in vitro assay to evaluate sperm penetration intocervical mucus. Bergman et al. (1981, Fertility and Sterility36:363-367) found excellent correlation between sperm penetration intofrozen bovine cervical mucus and fresh human cervical mucus (r=0.98) dueto the similarity of human and bovine cervical mucus rheological andbiophysical make-up (Bergman et al., Fertil. Steril. 36:363-367, 1981;Keel et al., Arch. Androl. 44:109-115, 2000). Human cervical mucus isproduced in small volumes, has variable quality in a clinical setting,and is difficult to access. In cattle, large quantities of estrus mucusare produced, making access straightforward. Additionally, cows areselected for high fertility, so cervical mucus quality is moreconsistent. Mucus from several cows can be pooled, frozen, andsubsequently used across an entire experiment, thereby decreasingvariability in the assay. De Geyter et al. (Hum. Reprod. 3:948-954,1988) found better correlation between human IVF outcomes and spermpenetration in bovine cervical mucus as compared to human cervicalmucus, using the endpoints of vanguard sperm penetration and spermdensity at fixed distances. Sharara et al. (1995, Human Reproduction10:1481-1485) found that sperm penetration into human or bovine cervicalmucus correlated with IVF outcomes at a similar rate (r=0.66). Keel andSchalue (Arch. Androl. 44:109-115, 2000) performed a large experimentwith 1,400 human ejaculates to observe the relationship betweenejaculate quality and bovine cervical mucus penetration test (BCMPT)outcomes. They found that sperm motility correlated well withpenetration in the BCMPT (r=0.448) in a linear relationship. However,approximately 30% of the samples with normal semen characteristics hadabnormal BCMPT. This indicated that the BCMPT was evaluating somethingunique that sperm motion characteristics alone could not detect (Shararaet al., Hum. Reprod. 10:1481-1485, 1995). Also, cryopreserved bull spermwith higher pregnancy rates in cows had a higher penetration rate intobovine cervical mucus than sperm from less fertile bulls (Tas et al.,Anim. Reprod. Sci. 101:18-27, 2007).

Alternatively, sperm penetration of mucus may be measured in vivo inwomen. At various times post-coitus, a sample of cervical mucus isremoved and examined microscopically for the number of sperm present inthe sample. In the post-coital test, improved sperm function isestablished if more sperm with faster velocity are seen in the mucussample after exposure to a composition of the present disclosure versusa sample of mucus from the patient after exposure to a controllubricant.

Other assays of sperm function potential include the sperm penetrationand hemizona assays. In the sperm penetration assay, the ability ofsperm to penetrate into an oocyte is measured. Briefly, commerciallyavailable zona free hamster oocytes are used (EmbryoTech Laboratories,Haverhill, Mass.). Hamster oocytes are suitable in this assay for spermof any species. Capacitated sperm, such as those cultured with bovineserum albumin for 18 hours, are incubated for 3 hours with the hamsteroocytes. Following incubation, oocytes are stained with acetolacmoid orequivalent stain and the number of sperm penetrating each oocyte iscounted microscopically. A hemizona assay measures the ability of spermto undergo capacitation and bind to an oocyte. Briefly, in this assay,live normal sperm are incubated in media with bovine serum albumin,which triggers capacitation. Sperm are then incubated with dead oocyteswhich are surrounded by the zona pellucida, an acellular coating ofoocytes. Capacitated sperm bind to the zona and the number of spermbinding is counted microscopically.

In certain embodiments, a composition of the present disclosure arenon-toxic to sperm if following exposure to a 10% solution of thecomposition, sperm retain at least 80% motility as compared to spermexposed to a control medium.

(2) Evaluating Oocyte or Embryo Quality

Compositions of the present disclosure may be evaluated for their effecton oocyte survival and function using a variety of methods known in theart. The effect of a test composition on oocyte quality may bedetermined using an oocyte expansion assay. The mammalian pre-ovulatoryfollicle contains a mature oocyte that is enclosed by the somaticcumulus cells forming the cumulus cell-oocyte complex (COC). Within thiscomplex, the cumulus cells are tightly connected to each other and tothe oocyte via cell adhesion complexes and gap junctions. In response tothe ovulatory luteinizing hormone (LH) surge, cumulus cells expand andproduce a complex extracellular matrix, which is essential forovulation, fertilization, and subsequent embryonic development. Thishighly coordinated process is called “cumulus expansion.” An in vitrocumulus expansion assay tests the ability of mouse oocytes to expandfollowing exposure to a composition. For example, mouse oocytes areincubated in medium with or without 50 μg/ml luteinizing hormone for 22hours. Normal oocytes will have >3-5 layers of expanded cumulus. Normalcumulus cell expansion is required for oocytes maturation andfertilization.

The effect of a test composition on oocyte quality may also be assessedby oocyte maturation status. Staining techniques for staging nuclearmaturation of oocytes include 1% aceto-orcein staining and anti-laminA/C and 4′,6-diamidino-2-phenylindole technique, which may be used todetermine the percentage of oocytes that progress to metaphase II stagefollowing exposure to a test composition. Oocytes entering metaphase IIstage have completed meiosis 1 to become a haploid oocyte and commencedmeiosis 2, arresting at metaphase until fertilization occurs. Onlyoocytes that are in the metaphase II stage are capable of beingfertilized.

The effect of a composition disclosed herein on embryonic quality may bedetermined using morphological evaluations in an animal model (e.g.,bovine, rodent), including observing for normal cleavage or divisionalof the embryo in culture (Lindner and Wright, Theriogenology 20:407,1983); normal formation of a blastocysts cavity at an appropriate timein culture; counting the number and health of cells found in the embryousing Hoeschst 33342 stain (Pursel et al., Theriogenology 24:687);establishment of pregnancy following transfer to a female subject; birthof a normal offspring following transfer to a female subject.

A mouse embryo assay (MEA) is a common system to assess toxicity ofmaterials used in assisted reproductive techniques. In brief, a one-cellembryo is obtained and exposed to a 10% solution of the test compositiondiluted in M2 mouse embryo culture media for a period of time (e.g., 30minutes). The embryos are then transferred to appropriate culture mediumand cultured for 96 hours. The number 1-cell embryos that transitionedto 2-cell embryos within 24 hours and the number of 1-cell embryos thattransitioned to expanded blastocyst stage within 96 hours (blastocystrate) are also determined. In certain embodiments, a test composition isnon-toxic to embryos if the blastocyst rate of the group exposed to thetest composition is at least 80% of the blastocyst rate of the controlgroup within 96 hours following exposure to the test composition.

(3) Evaluating Vaginal Flora

The vaginal microbiota (VMB) is a key component of regulating LRThomeostasis. Demonstration that a topical product administered to theLRT does not significantly harm Lactobacillus colonies and relatedbeneficial organisms is critical to maintaining or enhancing LRTfunction. To assess the impact of a topical composition on the VMB,testing of normal vaginal flora is performed using methods known in theart.

The effect of any composition disclosed herein on vaginal microbiota(e.g., Lactobacillus species) may be determined using the minimalmicrobicidal concentration method (MMC), the concentration required toreduce the viability of a culture by 99.99%, as described in Dezzutti etal. (PLoS One 7:e48328, 2012). Briefly, Lactobacillus suspensions areprepared by selecting isolated colonies from fresh overnight cultureplates and suspending the test organisms in saline to a cell density of2×10⁸ bacteria/ml. Test and control compositions are mixed with an equalvolume of the bacterial suspensions at room temperature and plated forcolony forming units (CFUs). The number of CFUs is taken from thedilution plate containing 50 to 300 colonies. Samples are taken at time0 and various time points post-exposure. Samples yielding 10 or fewerCFUs (representing a 99.99% kill) are considered sensitive to killing.All results are compared to the control (positive or negative), whichhave the same excipient vehicle but lack the active ingredients of thetest composition(s) or contain a known microbicide in the same excipientvehicle. In vitro methods of testing the effects of compositions onplanktonic Lactobacillus growth, vaginal epithelial cell viability,Lactobacillus-vaginal epithelial cell interactions have been describedin Fashemi et al. (2013, Microb. Ecol. Health Dis. 24:19703).

Other methods that may be used to assess changes in vaginal microbiotafollowing exposure to a LRT composition of this disclosure include 16SrRNA gene sequencing (see, e.g., Romero et al., Microbiome 2:4, 2014;Macklaim et al., Microb. Ecol. Health Dis. 26:27799, 2015), speciesspecific quantitative PCR (Zozaya-Hinchliffe et al., J. Clin. Microbiol.48:1812, 2010), and phylogenetic microarrays (Paliy and Agans, FEMSMicrobiol. Ecol. 79:2, 2012).

In certain embodiments, the compositions of the present disclosurereduce normal vaginal microbiota population (e.g., L. crispatus, L.gassseri, L. jensenii, L. acidophilus, or any combination thereof) by nomore than about 35%, about 30%, about 25%, about 20%, about 15%, about10%, or about 5% when exposed to a concentration of the composition thatis to be used in vivo on a female subject. In further embodiments,compositions of the present disclosure do not reduce normal vaginalmicrobiota of a female subject by more than about 35%, about 30%, about25%, about 20%, about 15%, about 10%, or about 5% when an effectiveamount of a composition of this disclosure is used to contact the LRT ofa female subject, wherein the amount of the composition used to contactthe LRT of a female subject causes less than a 1×log₁₀ reduction ingrowth of about one to about five Lactobacillus species in the minimalmicrobicidal concentration (MMC) assay of Dezzutti et al. (2012). Inother embodiments, compositions of the present disclosure do not reducethe vaginal microbiota of a female subject more than about 35%, about30%, about 25%, about 20%, about 15%, about 10%, or about 5% followingadministration of an effective amount of the composition to the lowerreproductive tract as compared to the vaginal microbiota of the femalesubject prior to administration of the composition, wherein a speciesspecific quantitative PCR measure of about one to about fiveLactobacillus species present in the vaginal microbiota of the subjectbefore and after contact with a composition of this disclosure isdetermined by the method of Zozaya-Hinchliffe et al. (2010).

(4) Irritation and Inflammation Assessment

The irritation and inflammation can be assessed in vitro or in vivousing human or animal vaginal-ectocervical tissue explants; vaginal,cervical or vulvar cell monolayers; skin cell monolayers; slug mucosalirritation assays; sperm toxicity assays; or other equivalent methods.

For example, the slug mucosal irritation assay (SMI) is a sensitivesystem to detect even mild mucosal irritation potency (Adriaens et al.,Sex. Transm. Dis. 35:512, 2008). The SMI assay uses slugs (Arionlusitanicus) as the test organism. The body wall of slugs consists of amucosal surface comprising mucus secreting cells covering asubepithelial connective tissue. Slugs that are placed on an irritantsubstance will actively produce mucus as a protective mechanism fromnoxious agents. Additionally, tissue damage of the slug's surfaceresults in the release of proteins and enzymes. The proteinconcentration in the collected samples is determined with a proteinquantitation kit. A composition of this disclosure is considerednon-irritating if it does not cause an increased production of mucus, oran increased release of proteins and enzymes as compared to a negativecontrol.

Human, organotypic vaginal-ectocervical tissue model produced fromnormal human-derived vaginal-ectocervical epithelial cells may also beused to assess irritation of topically applied products, as canmonolayers of cervical or vaginal epithelium (Ayehunie et al.,Toxicology 279:130, 2011; Ayehunie et al., 2007, Toxicology 279:130-8;Trifonova et al., 2006, Antimicrob. Agents Chemother. 50:4005-4010;Fichorova et al., 2011, mBio 2:e00168-11). Release of markers of celldamage (e.g., increase in CD4, IL-1β, CXCL8, CCL2, CCL21, EMP1; decreasein BPI) and production of inflammatory mediators, such as IL-1, IL-8,TLR4, may be used as markers of irritation and pro-inflammation (see,also Fichorova et al., 2015, Toxicol. Appl. Pharmacol. 285:198-206;Doncel et al., 2004, J. Acquir. Immune Defic. Syndr. 37(Suppl.3):S174-180; Fichorova et al., 2004, Biol. Reprod. 71:761-769; Moench etal., 2010, BMC Infect. Dis. 10:331). Biomarkers of epithelial integrityand immune function have been validated in multiple clinical studies ofvaginal product safety (Mauck et al., 2013, AIDS Res. Hum. Retroviruses29:1475-86; Fichorova et al., mBio, 2015, 6:e00221-15; Fichorova et al.,2011, Cytokine 55:134-140; Mauck et al., 2008, J. Acquir. Immune Defic.Syndr. 49:243-50; Morrison et al., 2014, J. Acquir. Immune Defic. Syndr.66:109-117; Schwartz et al., 2006, Contraception 74:133-40; Keller etal., 2003, J. Antimicr. Chemother. 51:1099-1102). A composition of thisdisclosure is considered non-irritating and non-inflammatory if it doesnot cause release of markers of cell damage or increased expression ofpro-inflammatory mediators above that are caused by a negative control(e.g., synthetic TLR2/6 ligand).

Compositions as disclosed herein may also be tested for their effects invaginal infection susceptibility models, such as a mouse genital herpestransmission model (see, e.g., Moench et al., 2010, BMC Infect. Dis.10:331). Increased susceptibility to infection transmission may becaused by vaginal epithelial cell toxicities due to exposure to toxicexcipients.

Effects of topical compositions on tissue viability using tissue models(e.g. human explants or cell monolayers) may also be assessed using theMTT colorimetric assay technique. The MTT assay is a colorimetric assayfor assessing cell metabolic activity. NAD(P)H-dependent cellularoxidoreductase enzymes may, under defined conditions, reflect the numberof viable cells present. These enzymes are capable of reducing thetetrazolium dye MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide to its insoluble formazan, which has a purple color. The MTTassay may be used to measure a composition's cytotoxicity or effect oncell viability (Ayehunie et al., 2011).

In addition, oxidative stress and antioxidant potential of the tissuescan be determined by common methods, such as a TBARS assay to evaluatethe impact of various embodiments on tissue health. Because reactiveoxygen species (ROS) have extremely short half-lives, they are difficultto measure directly. Instead, several products of the damage produced byoxidative stress, such as thiobarbituric acid reactive substances(TBARS), can be measured. TBARS are formed as a byproduct of lipidperoxidation (i.e., as degradation products of fat), which can bedetected by the TBARS assay using thiobarbituric acid as a reagent.

The in vivo rabbit vaginal irritation (RVI) model may also be used toassess the irritation and inflammatory characteristics of a formulation(Eckstein et al., J. Reprod. Fertil. 20:85, 1969). This model is basedon macroscopic observations of erythema, edema and ulceration andhistopathologic analysis of the tissues collected after exposure of theanimals to the test materials. A non-irritating and safe composition ofthis disclosure would show no negative macroscopic or histopathologiceffects as compared to a control vehicle. An expanded RVI model having aquantitative nuclease protection assay (qNPA) to quantify mRNA levels of25 genes representing leukocyte differentiation markers, toll-likereceptors (TLR), cytokines, chemokines, epithelial repair, microbicidaland vascular markers has also been described (see, Fichorova et al.,2015, Toxicol. Appl. Pharmacol. 285:198-206).

In another example, in vivo coloscopic exams of women following use ofLRT compositions can be used to identify signs of inflammation orirritation. User surveys can also be used for scoring of symptoms of thesame (Van Damme et al., Lancet 360:971, 2002; Bunge et al., J. Acquir.Immune Defic. Syndr. 60:337, 2012).

Sensitization tests may be used evaluate the potential of a compositionof this disclosure to cause a sensitizing effect or allergenic reactionin a female subject over an extended period of exposure. Exemplary testsinclude Guinea pig tests, such as the Magnusson-Kligman guinea pigmaximization test (J. Invest. Dermatol. 52:268, 1969), the occludedpatch test of Buehler (Arch. Dermatol. 91:171, 1965), and the openepicutaneous test (Kero et al., Contact Dermatitis 6:341, 1980).

In yet another example, a murine local lymph node assay (LLNA) isanother method for the identification of skin sensitizing chemicals. Incontrast to guinea pig tests, this assay relies on measurement of eventsinduced during the induction phase of skin sensitization, specificallylymphocyte proliferation in the draining lymph nodes which is a hallmarkof a skin sensitization response, rather than the elicitation phase(Kimber et al., Contact Dermatitis 21:215, 1989; Basketter et al., FoodChem. Toxicol. 34: 985, 1996).

The human repeat-insult patch test (HRIPT) may be performed as aconfirmatory test in the safety evaluation of skin sensitizers.Sensitization is a process by which humans develop increased allergicresponses to a substance over time through repeated exposure to thatsubstance. It is different from irritation because it involves an immuneresponse. Skin sensitization reactions are usually characterized byerythema coupled with one or more of various dermal sequelae, such asedema, papules, vesicles, bullae, and/or pruritis (McNamee et al.,Regul. Toxicol. Pharmacol. 52:24, 2008).

Irritating topical products may trigger the release of pro-inflammatorycytokines (e.g., TLR, IL-1, IL-6, IL-8, TNF-α, IFN-γ, IL-17) andinflammasomes (e.g., NLRP3 and NLRC4). Cytokines and inflammasomes canbe measured using an enzyme-linked immunosorbent assay (ELISA),quantitative PCR, or other molecular assay. A product is considerednon-inflammatory if it does not cause increased expression ofpro-inflammatory cytokines or inflammasomes (Rabeony et al., Eur. J.Immunol. 45:2847, 2015).

In certain embodiments, the compositions of the present disclosure donot induce irritation or inflammation potential in the lowerreproductive tract of a female subject greater than about 0.5%, about1%, about 2.5%, about 5%, about 10%, about 15%, about 20%, about 25%, orabout 30% as compared to an untreated control subject, preferably asmeasured using the slug test of Adriaens et al. (2008).

EXAMPLES Example 1 Effect on Sperm Viability of Isotonic Composition ofSalvia Extract

An isotonic, pH neutral (pH 7) solution comprising 10% Salvia hispanica(in oil format) was evaluated in a 24 hour sperm survival assay aspreviously described (see, Vargas et al., Fertil. Steril. 95:835, 2011).The isotonic solution comprising Salvia hispanica extract caused asignificant decrease in levels of lipid peroxidation (LPO) damage ascompared to that seen for sperm treated with control medium or withcanola oil or almond oil (see Table 3). Oxidative stress results inlipid peroxidation of the sperm membrane, resulting in membranedysfunction and impairment of sperm motility (Gil-Villa et al., Fertil.Sterol. 94:1465, 2010).

However, the isotonic solution comprising Salvia hispanica oilsurprisingly caused significant sperm toxicity over the 24 hr incubation(FIG. 1). A Sperm Toxicity Index of <0.75 (meaning treated sperm surviveon average less than 75% as compared to controls) is deemed ascytotoxic. The isotonic solution comprising Salvia hispanica oil exerteda profoundly significant (*p=0.0001) decrease in sperm motility over thesperm motility observed for control medium or with other natural plantproducts, i.e canola oil or almond oil, even though peroxidative damageto the sperm was mitigated with the Salvia (Table 3).

Taken together, these data suggest that Salvia hispanica has a toxiceffect on sperm that is separate from any cellular oxidative damage,which would also trigger inflammation in the female LRT. This spermtoxicity effect may be useful in embodiments aimed at supportinghomeostasis of the LRT during the non-ovulatory stage of a woman'scycle, when secretions are innately hostile to sperm function (e.g.,contraceptive compositions).

TABLE 3 Lipid Peroxidation Levels in Sperm Post-Treatment LPO productsTreatment (ng/ml) Control 168 Salvia hispanica Oil  103* Canola Oil 150Almond Oil 174

Example 2 Effect of Salvia sclarea Extract on Mucosal IrritationPotential

The mucosal irritation potential of various embodiments of isotonicsolutions that support LRT homeostasis was measured using a slug mucosalirritation (SMI) assay as described in Adriaens and Remon, 2008, Sex.Transm. Dis. 35:512-6. In brief, the SMI assay uses slugs (Arionlusitanicus) as a test organism. The body wall of slugs comprises amucosal surface including mucus secreting cells covering asub-epithelial connective tissue. Slugs that are placed on an irritantsubstance will actively produce mucus as a protective mechanism fromnoxious agents. Additionally, tissue damage of the slug's surfaceresults in the release of proteins and enzymes. The proteinconcentration in the collected samples is determined with a proteinquantitation kit. A product is considered non-irritating if it does notcause increase production of mucus, and release of proteins and enzymesabove that caused by a negative control. Fertility Gel 1a and FresheningGel 2a are isotonic gels containing xylose. These both caused “mild”irritation as measured by SMI. Unexpectedly, adding a Salvia sclareaextract to each gel formulation helped decrease mucus production to makea “non-irritating” embodiments for Fertility Gel Formula AB (alsoreferred to as Gel 1b) and Freshening Gel Formula AB (also referred toas Gel 2b) (FIG. 2).

Studies of Mucosal Irritation Potential in the Slug Mucosal Index (SMI),supports the methods and compositions disclosed herein for creatingisotonic, non-irritating products that support LRT homeostasis.

Example 3 pH Balancing and Freshening Gel

An embodiment for a pH balancing and freshening composition is providedin herein as Freshening Gel Formula AB (also referred to as Gel 2b).

Other freshening compositions, preferably formulated as a gel, thatcontain ingredients reported to be toxic to vaginal mucosal cells andvaginal microbiota caused moderate to severe irritation potential in theSMI (RepHresh and Monistat) (FIG. 3). Surprisingly, Freshening Gel 2bcaused no mucosal irritation potential (FIG. 3), even though theisotonic product is pH matched to the low acidity of the non-ovulatingvagina (i.e., pH 4.5). The control was an iso-osmotic, preservative-freehydroxyethylcellulose gel.

Unexpectedly, the other products tested caused a complete loss of spermmotility on contact as measured by the toxicology screen, sperm survivalassay (described in Vargas et al. Fertil. Steril., 2011, 95:835-6),whereas sperm motility was preserved on contact with the Gel 2b product(FIG. 4). The control in this experiment was buffered sperm culturemedium.

Gel 2b is assayed for detrimental effects on vaginal microbiota speciesas compared with RepHresh® gel and MONISTAT®. Lactobacillus species thatare indigenous to the healthy human vaginal microbiome (L. crispatus, L.gasseri, and L. jensenii) are incubated with Gel 2b, RepHresh®, orMONISTAT® to evaluate the effect of the gels on the vaginal microbes asdescribed in Fashemi et al., 2013, Microbial Ecology in Health & Disease24:19703. In brief, Lactobacillus crispata isolated from vaginal swabsamples from healthy women is used to prepare bacterial suspensions. Abacterial suspension is incubated with 10% concentration of testcomposition for 24 hours, serially diluted, and plated on agar plates toobtain colony forming units. Control sample is incubated with mediaonly. The gel formulations are also evaluated for effect on G.vaginalis, a bacterium that is implicated in the genesis of bacterialbiofilms and bacterial vaginosis. The test gels are then rankedaccording to impact on L. crispata cfu.

Freshening Gel 2b undergoes safety evaluation using a human organotypicmodel (Ayehunie et al., Toxicology 279:130, 2011; Klausner et al., 2007,Toxicology 279:130-8; Trifonova et al., 2006, Antimicrob. AgentsChemother. 50:4005-4010; Mauck et al., AIDS Res. Hum. Retroviruses,2013, 29:1475-1486). A three-dimensional organotypicvaginal-ectocervical (VEC) tissue model is constructed in vitro usingnormal VEC epithelial cells and is well stratified, containingdifferentiated basal, suprabasal, intermediate, and superficial celllayers similar to in vivo tissue. The organotypic model is assessedfollowing exposure to a test composition (Gel 2b, RepHresh®, orMONISTAT®) for signs of cytotoxicity or irritation as compared to acontrol sample comprising hydroxyethylcellulose). Biomarkers ofcytotoxicity or irritation include: (1) decreased epithelial viabilityover a period of 24 hours; (2) decreased epithelium-associatedLactobacillus colony-forming units; (3) increased epithelial disruptionbiomarkers (e.g., IL-1α, IL-1β); (4) increased pro-inflammatory markers(e.g., ratio of IL-1(α+β) to IL-1RA; IL-8).

Example 4 Endotoxin Level of D-Xylose

Analysis of plant derived products found an unexpected high level ofendotoxin in most Aloe Vera products. Many lubricants and other LRTproducts contain aloe vera, which could serve as an inflammatory triggerin LRT products. In contrast, xylose from a natural raspberry source(d-Xylose source 1) as used in the current embodiments has a very lowendotoxin level, supporting LRT homeostasis (Table 4).

TABLE 4 Endotoxin Level of D-Xylose Product Endotoxin Level Aloe VeraPowder source 1 209 EU/mg Aloe Vera Powder source 2 25 EU/mg D-Xylosesource 1 <0.07 EU/mg

Bacterial endotoxin levels in Fertility Gel 1b (Fertility CompositionFormula AB) and Freshening Gel 2b (Freshening Composition Formula AB)were assessed following the protocol set forth in the United StatesPharmacopeia and The National Formulary (USP-NF), Chapter 85. Bacterialendotoxin levels in Gel 1b and Gel 2b were demonstrated to be low (Table5).

TABLE 5 Endotoxin Level of Exemplary Fertility Gel 1b and Freshness Gel2b Test Gel Endotoxin Level Fertility Gel 1b <1 EU/ml Freshening Gel 2b<1 EU/ml

Example 5 Fertility Compositions

Products for enhanced sperm function and vaginal homeostasis during theovulatory stage of the cycle are disclosed. An isotonic, pH neutralFertility Gel 1b (Fertility Composition Formula AB) containing xyloseand Salvia sclarea extract resulted in significantly lower lipidperoxidation of sperm plasma membranes following the experimentdescribed in Gil-Villa et al. (Fertil. Steril. 94:1465, 2010) after 1hour of incubation as compared to a paraben-containing commercialfertility product containing methylparaben and glycerol (Table 6).Additionally, the Fertility Gel 1b (Fertility Composition Formula AB)formulation resulted in no mucosal irritation potential, whereas theparaben and glycerin-containing commercial fertility product caused someincrease in irritation potential, and surprisingly KY Jelly causedsevere irritation (FIG. 5).

TABLE 6 Lipid Peroxidation of sperm after 1 hr. incubation Fertility Gel1b 5.5 ng/ml +/− 1.0 Commercial Fertility Gel 7.5 ng/ml +/− 2.1Means differ at P=0.05

An additive antioxidant effect of Salvia embodiments is observed onsperm lipid peroxidation in culture after 3 hours. Fertility Gel 1a isan embodiment with xylose. Fertility Gel 1b (Fertility CompositionFormula AB) is an embodiment with xylose and salvia. Lipid peroxidationlevels were significantly lower when both xylose and salvia were present(Table 7, p≤0.005).

TABLE 7 Lipid Peroxidation of sperm after 3 hours incubation FertilityGel 1a 42 ng/ml +/− 6 Fertility Gel 1b 39 ng/ml +/− 8

Fertility Gel 1b (Fertility Composition Formula AB) was also assessedfor its effect on sperm motility and DNA damage (Table 8). Normal spermquality samples and low quality sperm samples (low sperm count) wereincubated for 30 minutes with a 10% solution of the Fertility Gel 1b orcontrol medium (human Fallopian tube fluid).

TABLE 8 Sperm Motility and DNA Damage after 30 min incubation with Gel1b Total % DNA Sample Sperm Quality Motile count Damage Control MediumNormal 43 +/− 17  9 +/− 5 Fertility Gel 1b Normal 43 +/− 16  9 +/− 4Control Medium Low Quality  5 +/− 1.5 11 +/− 4 Fertility Gel 1b LowQuality 5 +/− 1 10 +/− 3

The effect of Fertility Gel 1b on the ability of cryopreserved bullsperm to penetrate estrus cow cervical mucus following 30 minuteexposure to a 10% solution was evaluated using a cervical mucuspenetration assay (Table 9).

TABLE 9 Bovine Cervical Mucus Penetration Assay Vanguard Sperm # Spermat Sample Distance 3 cm Control Medium (TALP) 5.5 cm +/− 1 12 +/− 3 10%Fertility Gel 1b 5.8 cm +/− 2 16 +/− 2

Means differ between control and treated for both Vanguard sperm andsperm numbers at 3 cm (paired t-test, p≤0.05)

Taken together, Fertility Gel 1b provides: sperm antioxidant supportwith less oxidative damage to post-ejaculatory sperm, decreased mucosalirritation potential, superior cervical mucus penetration, and isnon-detrimental to sperm motility or DNA quality.

Fertility Gel 1b (Fertility Composition Formula AB) was also evaluatedfor its effect on embryonic quality using a 1-cell mouse embryonic assay(MEA). The MEA is used as a regulatory special control for class IIassisted reproduction and fertility devices and is a surrogate indicatorof potential toxicity of materials used in fertility interventions thatcontact gametes and/or embryos (Gardner et al. 2005, Seminars in Reprod.Med. 23:319-324; Van den Bergh et al., 1996, J. Assisted Reprod. Genet.13:733-738). In brief, one-cell mouse embryos were obtained and exposedto a 10% solution of the test composition diluted in M2 mouse embryoculture media for 30 minutes. The embryos were then transferred toappropriate culture medium and cultured for 96 hours. The number 1-cellembryos that transitioned to 2-cell embryos within 24 hours and thenumber of 1-cell embryos that transitioned to expanded blastocyst stagewithin 96 hours (blastocyst rate) were also determined. As shown inTable 10, Fertility Gel 1b test composition is non-toxic to embryos witha blastocyst rate that is greater than 80% of the control group.

TABLE 10 Mouse Embryo Assay for Fertility Gel 1b # 2-Cell EmbryosBlastocyst Rate Sample at 24 hrs at 96 hrs Gel 1b 15/15 15/15 Control20/21 21/21

An isotonic, pH neutral Fertility Gel 1c (also referred to herein asFertility Gel Formula AB′) was assessed for its effect on human spermmotility. The effect of a 10% solution of Fertility Gel 1c on spermmotility parameters was determined on semen collected from healthy,human male donors with both normal sperm counts (normospermia, n=10) andlow total motile sperm counts (oligoasthenozoospermic, n=10) from alocal sperm bank. Normospermia was defined as total motile sperm countsof >20 million/ml and oligoasthenozoospermia (OAT) was defined as totalmotile sperm concentrations of <15 million/ml. Each specimen wasproduced by masturbation without lubricant into a sterile plasticcontainer after a recommended abstinence period of 48-96 hours.Specimens were allowed to liquefy and then processed within 30 minutes.Each sperm sample was diluted 1:4 with HTF media+10% human serumalbumin, divided into two 900 μl aliquots, and placed into tissueculture wells. Gel 1c was added to one aliquot to achieve finalconcentrations of 10% V/V. The second aliquot served as the control.Specimens were incubated for 30 minutes at 37° C. and 5% CO₂ in 95%humidity.

The percentages of progressively motile sperm were determined manuallyin the following manner: 10 μl samples were placed on pre-warmed slides,and covered with a coverslip. Evaluations were performed according toWHO guidelines using an inverted phase contrast microscope.Progressively motile sperm were those regarded as a+b forms as indicatedby WHO guidelines. Total motile concentration was calculated as theproduct of total sperm count and percent progressively motile spermdivided by 100. Normospermic and OAT semen samples exposed to 10% Gel 1cfor 30 minutes did not differ from control with respect to percentmotility (FIG. 6) or total motile concentration (FIG. 7). Thus, fornormal and OAT samples, Gel 1c treatment did not harm sperm motility andtotal motile sperm concentrations.

The impact of Fertility Gel 1b (Fertility Formula AB) on sperm geneticmaterial was assessed using a human sperm chromatin structure assay(SCSA). This assay is based on the metachromatic staining of single anddouble stranded DNA by acridine orange stain (Evenson et al., 1999,Human Reprod. 14:1039-1049; Evenson et al., 2002, J. Andrology23:25-43). Excitation with an argon laser causes acridine orangeintercalated in double-stranded DNA to emit a green fluorescence,whereas red fluorescence is emitted by single strand DNA. The extent ofDNA denaturation in a sample is expressed as a and calculated by theformula α=red/(red+green). The endpoint measurement is DNA FragmentationIndex (DFI).

Semen samples were obtained from healthy male donors with normospermia(n=5) and mild oligoasthenozoospermia (n=5) at a local sperm bank.Normospermia was defined as total motile count>20 million/ml andoligoasthenozoospermia (OAT) was defined as total motile spermconcentration<15 million/ml. Each specimen was produced by masturbationwithout Gel 1b lubricant into a sterile plastic container after arecommended abstinence period of 48-96 hours. Specimens were allowed toliquefy and then processed within 30 minutes. Specimens were allowed toliquefy and then processed within 30 minutes. Specimens were allowed toliquefy and then processed within 30 minutes. Each sperm sample wasdiluted 1:4 with HTF media+10% human serum albumin, divided into two 900μl aliquots, and placed into tissue culture wells. Gel 1b was added toone aliquot to achieve final concentrations of 10% V/V. The secondaliquot served as the control. Specimens were incubated for 30 minutesat 37° C. and 5% CO₂ in 95% humidity. After 30 minutes of incubation,portions of each treatment were removed and placed in vials for flashfreezing in liquid nitrogen. These were then tested using the SpermChromatin Structure Assay as described.

The mean DFI (+/−sd) for normospermic, and OAT samples, was under 15%and did not differ between treated and control sperm (FIG. 8). A DFI of<15% DFI indicates excellent to good sperm DNA integrity. In normal andOAT human sperm samples, Fertility Gel 1b had no detrimental effect onDNA fragmentation or human sperm chromatin quality.

Fertility Gel 1c (Fertility Formula AB′) was assessed via the BovineCervical Mucus Penetration Test to determine whether it had anydetrimental effect on sperm penetration into cervical mucus. Cervicalmucus was collected and pooled from 10 cows. Cervical mucus was frozenin aliquots of 3 mls and stored at −20° C. until use. Frozen mucus wasthawed at room temperature for 30 minutes before use. Commercialcryopreserved sperm from 2 bulls was pooled and used in all replicates.Semen straws (0.5 mL) were thawed and sperm concentration adjusted to 10million spermatozoa per ml using a commercial bovine semen-freezingmedium. Flattened capillary tubes (0.3×3.0×100 mm) were filled withcervical mucus, sealed at one end and placed in 0.5 mL of the spermsuspension in each of two treatments: (1) Control (extender alone,lubricant free), and (2) Extender with 10% Gel 1c. Each treatment wasrepeated 10 times. Parameters evaluated after incubation at 37° C. for30 minutes were: (1) distance travelled by vanguard sperm (cm); (2)density of spermatozoa at 2 cm and 3 cm; and (3) percent progressivelymotile sperm at 30 minutes. Table 11 summarizes the results of thebovine cervical mucus penetration test. Sperm treated with Fertility Gel1c showed significantly higher rates of penetration into 3 cm of themucus columns than did the control sperm (p=0.0261).

TABLE 11 Mean ± SEM for cervical mucus penetration variables accordingto treatment Progressive Vanguard Motility sperm Density at Density atTreatment N (%) (cm) 2 cm 3 cm Control 10 40.5 ± 2.38 5.64 ± 0.20 27.9 ±1.10 14.4 ± 0.87  Gel 1c 10 42.0 ± 1.53 5.81 ± 0.17 29.7 ± 0.56 16.6 ±0.64* *p < 0.05 compared to control

The Fertility Gel 1c did not interfere with sperm motility in thissystem. Fertility Gel 1c also did not inhibit cervical mucus penetrationas determined by both the vanguard sperm distance, and sperm density atset distances in cervical mucus columns incubated with bull sperm.Indeed, Fertility Gel 1c significantly improved sperm penetration intothe furthest column distance (p<0.05).

Example 6 Slipperiness of Fertility Gel

A focus group was used to obtain quantitative measures of the degree of“lubricity” or “slipperiness” perceived for the Fertility Gel 1c(Fertility Formula AB′) versus a commercially available,paraben-containing fertility product.

The two gel products were applied onto two different 4-cm circular siteson the non-dominant forearms of participants (n=20). The order of thegel, and the circular site to which it was applied, were determinedrandomly based on a computer generated randomization table. Eachtreatment was replicated twice.

Fingers and circular sites were wiped with a baby-wipe and paper towel.The focus group leader dispensed 0.1 ml of each sample into the centerof the circular site previously drawn on the forearm. The sample wasrubbed in a circle using the index finger at a rate of 2 rps for 15seconds and followed with another 45 seconds. A metronome was set at 120bpm and played to standardize the manipulation rate.

At time points 15 seconds and 60 seconds, slipperiness was evaluatedusing a 10 cm visual analog scale (VAS) (FIG. 9A). The two ends of thevisual analog scale were anchored by the phrases “not slippery” and“very slippery.” The focus group participant was asked to make a singleline perpendicular to the VAS at the point of their slipperinessassessment between the two anchors. The degree of slipperiness wasdetermined by measuring the distance (cm) of the participants mark onthe visual analog scale from the left (“Not slippery”) baseline.

FIG. 9B demonstrates that the Fertility Gel 1c was at least as slipperyor lubricious as the commercial fertility gel at both 15 seconds (7.4 cmvs 7.2 cm, respectively) and 60 seconds (5.7 cm vs 5.4 cm,respectively). Thus, Fertility Gel 1c was as lubricious as a commercialparaben-containing fertility product.

Example 7 Menopause Lubricant Gels

Compositions directed at alleviating post-menopausal vaginal dryness anddiscomfort are described.

Menopausal Formula AB Gel was evaluated for irritation potentialcompared with other commercially available lubricants using the slugmucosal irritation (SMI) assay as described in Example 2. As shown inTable 12, Menopausal Formula AB Gel showed no irritation potential ortissue damage.

TABLE 12 SMI Assay for Menopausal Formula AB Gel Irritation TissueSample Potential Damage Menopausal Formula AB None No Gel KY JellySevere No Replens Moderate No Astroglide Severe Yes

Menopausal Formula AB Gel is assayed for detrimental effects onsuspensions vaginal microbiota species as described in Example 3.

The various embodiments described above can be combined to providefurther embodiments. All of the U.S. patents, U.S. patent applicationpublications, U.S. patent applications, foreign patents, foreign patentapplications and non-patent publications referred to in thisspecification and/or listed in the Application Data Sheet, including butnot limited to U.S. A No. 62/323,516 filed Apr. 15, 2016 and U.S.application Ser. No. 15/207,307 filed Jul. 11, 2016, are incorporatedherein by reference, in their entirety. Aspects of embodiments can bemodified, if necessary, to employ concepts of various patents,applications and publications to provide further embodiments.

These and other changes can be made to the embodiments in light of theabove-detailed description. In general, in the following claims, theterms used should not be construed to limit the claims to the specificembodiments disclosed in the specification and the claims, but should beconstrued to include all possible embodiments along with the full scopeof equivalents to which such claims are entitled. Accordingly, theclaims are not limited by the disclosure.

What is claimed is:
 1. A topical, isotonic vaginal gel composition foruse in a post-menopausal female subject, comprising: (a) xylose at atotal concentration ranging from about 0.01% to about 1% by weight; (b)a Salvia extract at a total concentration ranging from about 0.01% toabout 1% by weight; (c) a non-irritating viscosity increasing agent at atotal concentration ranging from about 0.1% to about 10% by weight; (d)a pH modifying agent at a total concentration ranging from about 0.001%to about 2% by weight; (e) a buffering agent at a total concentrationranging from about 0.5% to about 4% by weight; an osmolality adjuster ata total concentration ranging from about 0.1% to about 1% by weight; and(g) water at a total concentration ranging from about 85% to about 98%by weight; wherein the composition has a pH ranging from about 4.5 toabout 6.8.
 2. The composition of claim 1, wherein the non-irritatingviscosity increasing agent is a cellulose ester, a carbomer, apolyoxazoline, or any combination thereof.
 3. The composition of claim1, wherein the non-irritating viscosity increasing agent is ahypromellose, carbomer, carboxymethylcellulose,poly(2-methyl-2-oxazoline), or any combination thereof.
 4. Thecomposition of claim 1, wherein the non-irritating viscosity increasingagent is hypromellose and carbomer homopolymer type B together at atotal concentration ranging from about 0.1% to about 5%, wherein theratio of hypromellose to carbomer homopolymer type B in the compositionranges from about 2:1 to about 8:1.
 5. The composition of claim 1,wherein the buffering agent is sodium phosphate, potassium phosphate,citric acid, sodium citrate, or any combination thereof.
 6. Thecomposition of claim 1, wherein the buffering agent is monobasic sodiumand dibasic potassium phosphate at a total concentration ranging fromabout 0.5% to about 1.0% by weight.
 7. The composition of claim 1,wherein the pH modifying agent is ammonia, ammonium carbonate,diethanolamine, monoethanolamine, potassium hydroxide, potassiumphosphate dibasic, sodium bicarbonate, sodium borate, sodium carbonate,sodium hydroxide, sodium lactate, sodium phosphate dibasic, trolamine,lactic acid, or any combination thereof.
 8. The composition of claim 1,wherein the pH modifying agent is sodium hydroxide at a totalconcentration ranging from about 0.01% to about 1% by weight.
 9. Thecomposition of claim 1, wherein the osmolality adjuster is sodiumchloride, potassium chloride, or both.
 10. The composition of claim 1,wherein the Salvia extract is obtained from Salvia sclarea, Salviahispanica, Salvia plebeian, or any combination thereof.
 11. Thecomposition of claim 1, further comprising a paraben-free preservativeat a total concentration ranging from about 0.005% to about 5% byweight.
 12. The composition of claim 11, wherein the paraben-freepreservative is a phenethyl alcohol, caprylyl glycol, phenoxyethanol,benzoic acid, a benzoate, a sorbate, cranberry extract, oleuropein,acemannan, carvacrol, or any combination thereof.
 13. The composition ofclaim 1, further comprising a surfactant at a total concentrationranging from about 0.5% to about 5% by weight.
 14. The composition ofclaim 1, wherein: the Salvia extract of the composition comprises aSalvia sclarea extract at a total concentration ranging from about0.025% to about 1% by weight; the non-irritating viscosity increasingagent comprises hypromellose and carbomer homopolymer type B together ata total concentration ranging from about 1% to about 3% by weight andthe ratio of hypromellose to carbomer homopolymer type B in thecomposition is about 6.2:1; the buffering agent comprises a dibasicpotassium phosphate and a monobasic sodium phosphate at a totalconcentration ranging from about 0.5% to about 1% by weight; theosmolality adjuster comprises sodium chloride at a total concentrationranging from about 0.1% to about 1% by weight; the water is at aconcentration ranging from about 90% to about 98% by weight; and the pHmodifier comprises sodium hydroxide and lactic acid at a totalconcentration ranging from about 0.01% to about 1% by weight.
 15. Thecomposition of claim 14, wherein the surfactant is a cetylhydroxyethylcellulose, polyvinyl alcohol, poloxamer, or any combinationthereof.
 16. The composition of claim 14, further comprising aparaben-free preservative that comprises a solution of phenethyl alcoholand caprylyl glycol together at a total concentration ranging from about0.1% to about 1% by weight, wherein the solution comprises phenethylalcohol in an amount of 56%-60% and caprylyl glycol in an amount of40%-44%.
 17. The composition of claim 1, wherein: the Salvia extract ofthe composition comprises a Salvia sclarea extract at a totalconcentration ranging from about 0.025% to about 0.075% by weight; thenon-irritating viscosity increasing agent comprises hypromellose at atotal concentration ranging from about 0.75% to about 1.25% by weight;the buffering agent comprises a dibasic sodium phosphate and a monobasicsodium phosphate at a total concentration ranging from about 0.5% toabout 1% by weight; the osmolality adjuster comprises sodium chloride ata total concentration ranging from about 0.1% to about 0.7% by weight;the water is at a concentration ranging from about 95% to about 98% byweight; the pH modifier comprises sodium hydroxide at a concentrationranging from about 0.001% to about 1% by weight; and wherein thecomposition further comprises a surfactant, the surfactant comprisingpolyvinyl alcohol and poloxamer together at a total concentrationranging from about 2% to about 2.5% by weight.
 18. The composition ofclaim 17, further comprising a paraben-free preservative that comprisesbenzoic acid at a total concentration ranging from about 0.05% to about0.25% by weight.
 19. The composition of claim 1, wherein: the Salviaextract of the composition comprises a Salvia sclarea extract at a totalconcentration ranging from about 0.025% to about 0.075% by weight; thenon-irritating viscosity increasing agent comprises hypromellose at atotal concentration ranging from about 1.25% to about 2.25% by weight;the buffering agent comprises a dibasic sodium phosphate and a monobasicsodium phosphate at a total concentration ranging from about 0.5% toabout 1% by weight; the osmolality adjuster comprises sodium chloride ata total concentration ranging from about 0.1% to about 0.7% by weight;the water is at a concentration ranging from about 95% to about 98% byweight; and the pH modifier comprises sodium hydroxide at aconcentration ranging from about 0.001% to about 1% by weight; andwherein the composition further comprises a surfactant, the surfactantcomprising poloxamer at a total concentration ranging from about 0.75%to about 1.25% by weight.
 20. The composition of claim 19, furthercomprising a paraben-free preservative that comprises cranberry extractat a total concentration ranging from about 0.01% to about 0.05% byweight.
 21. The composition of claim 1, wherein: the Salvia extract ofthe composition comprises a Salvia plebeia extract at a totalconcentration ranging from about 0.025% to about 0.75% by weight; thenon-irritating viscosity increasing agent comprises carbomer homopolymertype B and hypromellose at a total concentration ranging from about0.10% to about 1% by weight and the ratio of hypromellose to carbomerhomopolymer type B in the composition is about 3:1; the buffering agentcomprises a monobasic sodium phosphate and dibasic sodium phosphate at atotal concentration ranging from about 0.5% to about 1% by weight; theosmolality adjuster comprises sodium chloride at a total concentrationranging from about 0.1% to about 0.7% by weight; the water is at aconcentration ranging from about 95% to about 99% by weight; and the pHmodifier comprises sodium hydroxide at a concentration ranging fromabout 0.001% to about 1% by weight.
 22. The composition of claim 21,further comprising a paraben-free preservative that comprises phenethylalcohol and caprylyl glycol together at a total concentration rangingfrom about 0.25% to about 0.5% by weight.
 23. The composition of claim1, wherein: the Salvia extract of the composition comprises a Salviasclarea extract at a total concentration ranging from about 0.025% toabout 0.075% by weight; the non-irritating viscosity increasing agentcomprises hypromellose and carboxymethyl cellulose together at a totalconcentration ranging from about 1.5% to about 2.5% by weight and theratio of hypromellose to carboxymethyl cellulose in the composition isabout 1.3:1; the buffering agent comprises citric acid and sodiumcitrate together at a total concentration ranging from about 1% to about1.25% by weight; the osmolality adjuster comprises sodium chloride at atotal concentration ranging from about 0.1% to about 0.7% by weight; thewater is at a concentration ranging from about 95% to about 98% byweight; and the pH modifier comprises sodium hydroxide at aconcentration ranging from about 0.001% to about 1% by weight.
 24. Thecomposition of claim 23, further comprising a paraben-free preservativethat comprises potassium sorbate and sodium benzoate together at a totalconcentration ranging from about 0.25% to about 0.5% by weight.
 25. Amethod of lubricating the lower reproductive tract of a post-menopausalfemale subject, comprising topically applying an effective amount of thecomposition of claim 1 to the lower reproductive tract of thepost-menopausal female subject.
 26. The method of claim 25, wherein thevaginal microbiota of the post-menopausal female subject is not reducedgreater than 25% following topical application of the composition to thelower reproductive tract, as compared to an untreated controlpost-menopausal female subject or prior to topical application of thecomposition to the lower reproductive tract.
 27. The method of claim 26,wherein the post-menopausal female subject is human.